From bbdfc4f8f5460d9860d5318e8bcbdc85a776442b Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Thu, 3 Aug 2023 13:18:31 +0000 Subject: [PATCH 001/123] The ".sandbox" pseudo-project is deprecated --- assets/samplesheet_test_full.csv | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/samplesheet_test_full.csv b/assets/samplesheet_test_full.csv index 599c5b7..4495dff 100644 --- a/assets/samplesheet_test_full.csv +++ b/assets/samplesheet_test_full.csv @@ -1,2 +1,2 @@ sample,datatype,datafile,indexfile -icCanRufa1,pacbio,/lustre/scratch123/tol/projects/.sandbox/data/insects/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram,/lustre/scratch123/tol/projects/.sandbox/data/insects/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram.crai +icCanRufa1,pacbio,/lustre/scratch123/tol/resources/nextflow/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram,/lustre/scratch123/tol/resources/nextflow/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram.crai From 683652fec79fdfdba94b38e864b2e3ae0d8372e0 Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Thu, 3 Aug 2023 13:18:39 +0000 Subject: [PATCH 002/123] Fixed a template error --- assets/slackreport.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/slackreport.json b/assets/slackreport.json index 043d02f..837b798 100644 --- a/assets/slackreport.json +++ b/assets/slackreport.json @@ -3,7 +3,7 @@ { "fallback": "Plain-text summary of the attachment.", "color": "<% if (success) { %>good<% } else { %>danger<%} %>", - "author_name": "sanger-tol/readmapping v${version} - ${runName}", + "author_name": "sanger-tol/variantcalling v${version} - ${runName}", "author_icon": "https://www.nextflow.io/docs/latest/_static/favicon.ico", "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors<% } %>", "fields": [ From a3eb2176660cf2c64734eaa3f8f87c090d21939e Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Fri, 4 Aug 2023 17:44:56 +0000 Subject: [PATCH 003/123] Our new convention is use per-pipeline directories under pipeline_info/ --- conf/modules.config | 4 ++-- docs/output.md | 2 +- lib/NfcoreTemplate.groovy | 2 +- nextflow.config | 2 +- nextflow_schema.json | 2 +- 5 files changed, 6 insertions(+), 6 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 0f4e51c..7dc8677 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -14,7 +14,7 @@ process { withName: SAMPLESHEET_CHECK { publishDir = [ - path: { "${params.outdir}/variantcalling_info" }, + path: { "${params.outdir}/pipeline_info/variantcalling" }, mode: params.publish_dir_mode, saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] @@ -48,7 +48,7 @@ process { withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ - path: { "${params.outdir}/variantcalling_info" }, + path: { "${params.outdir}/pipeline_info/variantcalling" }, mode: params.publish_dir_mode, pattern: '*_versions.yml' ] diff --git a/docs/output.md b/docs/output.md index 946a1c1..cfadaac 100644 --- a/docs/output.md +++ b/docs/output.md @@ -31,7 +31,7 @@ The aligned PacBio read data is used to call variants with DeepVariant. This is
Output files -- `variantcalling_info/` +- `pipeline_info/variantcalling/` - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. - Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`. diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 79cfdb4..bbd2fc1 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -120,7 +120,7 @@ class NfcoreTemplate { } // Write summary e-mail HTML to a file - def output_d = new File("${params.outdir}/pipeline_info/") + def output_d = new File("${params.outdir}/pipeline_info/variantcalling") if (!output_d.exists()) { output_d.mkdirs() } diff --git a/nextflow.config b/nextflow.config index cb204b4..f0e7b56 100644 --- a/nextflow.config +++ b/nextflow.config @@ -20,7 +20,7 @@ params { // Boilerplate options outdir = 'results' - tracedir = "${params.outdir}/variantcalling_info" + tracedir = "${params.outdir}/pipeline_info/variantcalling" publish_dir_mode = 'copy' email = null email_on_fail = null diff --git a/nextflow_schema.json b/nextflow_schema.json index c3e2d8b..7baf0c4 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -209,7 +209,7 @@ "tracedir": { "type": "string", "description": "Directory to keep pipeline Nextflow logs and reports.", - "default": "${params.outdir}/pipeline_info", + "default": "${params.outdir}/pipeline_info/variantcalling", "fa_icon": "fas fa-cogs", "hidden": true }, From 3fc2c953ff4460d687860cbebe6628c953268214 Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Sun, 13 Aug 2023 23:03:18 +0000 Subject: [PATCH 004/123] Latest nomenclature --- .github/workflows/{sangertest.yml => sanger_test.yml} | 0 .github/workflows/{sangerfulltest.yml => sanger_test_full.yml} | 0 2 files changed, 0 insertions(+), 0 deletions(-) rename .github/workflows/{sangertest.yml => sanger_test.yml} (100%) rename .github/workflows/{sangerfulltest.yml => sanger_test_full.yml} (100%) diff --git a/.github/workflows/sangertest.yml b/.github/workflows/sanger_test.yml similarity index 100% rename from .github/workflows/sangertest.yml rename to .github/workflows/sanger_test.yml diff --git a/.github/workflows/sangerfulltest.yml b/.github/workflows/sanger_test_full.yml similarity index 100% rename from .github/workflows/sangerfulltest.yml rename to .github/workflows/sanger_test_full.yml From 571735a41985b922f8fd59a07e1bb56da6c2fcdc Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Mon, 14 Aug 2023 13:42:51 +0000 Subject: [PATCH 005/123] Use the latest version of the Tower action --- .github/workflows/sanger_test.yml | 2 +- .github/workflows/sanger_test_full.yml | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index 7e8af27..6f69ddf 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -9,7 +9,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: nf-core/tower-action@v2 + uses: seqeralabs/action-tower-launch@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} diff --git a/.github/workflows/sanger_test_full.yml b/.github/workflows/sanger_test_full.yml index 850d4ae..fc11b07 100644 --- a/.github/workflows/sanger_test_full.yml +++ b/.github/workflows/sanger_test_full.yml @@ -22,7 +22,7 @@ jobs: if: github.event_name == 'workflow_dispatch' - name: Launch workflow via tower - uses: nf-core/tower-action@v2 + uses: seqeralabs/action-tower-launch@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} From c8483bab4ea58804fe86d66646cf4dbee2cd7b90 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 11:19:17 +0100 Subject: [PATCH 006/123] Wrong version number and keep the tower log files --- .github/workflows/sanger_test.yml | 9 ++++++++- .github/workflows/sanger_test_full.yml | 9 ++++++++- 2 files changed, 16 insertions(+), 2 deletions(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index 6f69ddf..db18591 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -9,7 +9,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@v2 + uses: seqeralabs/action-tower-launch@2.0.0 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} @@ -22,3 +22,10 @@ jobs: "outdir": "${{ secrets.TOWER_WORKDIR_PARENT }}/results/${{ github.repository }}/results-${{ github.sha }}", } profiles: test,sanger,singularity,cleanup + + - uses: actions/upload-artifact@v3 + with: + name: Tower debug log file + path: | + tower_action_*.log + tower_action_*.json diff --git a/.github/workflows/sanger_test_full.yml b/.github/workflows/sanger_test_full.yml index fc11b07..0bce7ee 100644 --- a/.github/workflows/sanger_test_full.yml +++ b/.github/workflows/sanger_test_full.yml @@ -22,7 +22,7 @@ jobs: if: github.event_name == 'workflow_dispatch' - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@v2 + uses: seqeralabs/action-tower-launch@v2.0.0 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} @@ -35,3 +35,10 @@ jobs: "outdir": "${{ secrets.TOWER_WORKDIR_PARENT }}/results/${{ github.repository }}/results-${{ env.REVISION }}", } profiles: test_full,sanger,singularity,cleanup + + - uses: actions/upload-artifact@v3 + with: + name: Tower debug log file + path: | + tower_action_*.log + tower_action_*.json From e876f6a3182eaf6c6426d6d0783e6b50544f87d7 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 11:46:48 +0100 Subject: [PATCH 007/123] just a test --- .github/workflows/sanger_test.yml | 3 +++ 1 file changed, 3 insertions(+) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index db18591..5021082 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -2,6 +2,9 @@ name: sanger-tol LSF tests on: workflow_dispatch: + push: + branches: + - tower_action jobs: run-tower: From 4034b3ed237e32f7aa074e062203cc9995348824 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 11:48:42 +0100 Subject: [PATCH 008/123] v2.0.0 doesn't work --- .github/workflows/sanger_test.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index 5021082..0f5ea38 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -12,7 +12,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@2.0.0 + uses: seqeralabs/action-tower-launch@2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} From 808e07376330c03726e34ddc53cf2abce2f256fd Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 11:50:16 +0100 Subject: [PATCH 009/123] using v2 --- .github/workflows/sanger_test.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index 0f5ea38..c956f2d 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -12,7 +12,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@2 + uses: seqeralabs/action-tower-launch@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} From 5f9589ec6263142e57b3e441d4abb2f9876ae467 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 21:43:36 +0100 Subject: [PATCH 010/123] delete the pipeline variable, using the default one - which will be the full url of repo --- .github/workflows/sanger_test.yml | 1 - 1 file changed, 1 deletion(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index c956f2d..e8234c6 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -17,7 +17,6 @@ jobs: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - pipeline: ${{ github.repository }} revision: ${{ github.sha }} workdir: ${{ secrets.TOWER_WORKDIR_PARENT }}/work/${{ github.repository }}/work-${{ github.sha }} parameters: | From 6b23886f7e611ee6790357cdb0dfb7fe7eb45091 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 21:47:00 +0100 Subject: [PATCH 011/123] finish the test and put everything back to normal --- .github/workflows/sanger_test.yml | 3 --- .github/workflows/sanger_test_full.yml | 1 - 2 files changed, 4 deletions(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index e8234c6..28f7625 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -2,9 +2,6 @@ name: sanger-tol LSF tests on: workflow_dispatch: - push: - branches: - - tower_action jobs: run-tower: diff --git a/.github/workflows/sanger_test_full.yml b/.github/workflows/sanger_test_full.yml index 0bce7ee..e0d5bc5 100644 --- a/.github/workflows/sanger_test_full.yml +++ b/.github/workflows/sanger_test_full.yml @@ -27,7 +27,6 @@ jobs: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - pipeline: ${{ github.repository }} revision: ${{ env.REVISION }} workdir: ${{ secrets.TOWER_WORKDIR_PARENT }}/work/${{ github.repository }}/work-${{ env.REVISION }} parameters: | From 8a7db40723fd2eddc75267500a246fb90db759a9 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 15 Aug 2023 21:53:59 +0100 Subject: [PATCH 012/123] put back the version number as v2 --- .github/workflows/sanger_test_full.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/sanger_test_full.yml b/.github/workflows/sanger_test_full.yml index e0d5bc5..e028c6b 100644 --- a/.github/workflows/sanger_test_full.yml +++ b/.github/workflows/sanger_test_full.yml @@ -22,7 +22,7 @@ jobs: if: github.event_name == 'workflow_dispatch' - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@v2.0.0 + uses: seqeralabs/action-tower-launch@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} From ebc3b229cb331f43a368a9c87f1fedd9c6fc0d7f Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Thu, 17 Aug 2023 16:37:55 +0000 Subject: [PATCH 013/123] Use the tolsoft access token to fix linting --- .github/workflows/fix-linting.yml | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml index 063af6e..dfb9b87 100644 --- a/.github/workflows/fix-linting.yml +++ b/.github/workflows/fix-linting.yml @@ -8,21 +8,21 @@ jobs: # Only run if comment is on a PR with the main repo, and if it contains the magic keywords if: > contains(github.event.comment.html_url, '/pull/') && - contains(github.event.comment.body, '@nf-core-bot fix linting') && + contains(github.event.comment.body, '@sanger-tol-bot fix linting') && github.repository == 'sanger-tol/variantcalling' runs-on: ubuntu-latest steps: - # Use the @nf-core-bot token to check out so we can push later + # Use the @sanger-tol-bot token to check out so we can push later - uses: actions/checkout@v3 with: - token: ${{ secrets.nf_core_bot_auth_token }} + token: ${{ secrets.sangertol_bot_access_token }} # Action runs on the issue comment, so we don't get the PR by default # Use the gh cli to check out the PR - name: Checkout Pull Request run: gh pr checkout ${{ github.event.issue.number }} env: - GITHUB_TOKEN: ${{ secrets.nf_core_bot_auth_token }} + GITHUB_TOKEN: ${{ secrets.sangertol_bot_access_token }} - uses: actions/setup-node@v3 @@ -46,8 +46,8 @@ jobs: - name: Commit & push changes if: steps.prettier_status.outputs.result == 'fail' run: | - git config user.email "core@nf-co.re" - git config user.name "nf-core-bot" + git config user.email "tolsoft@sanger.ac.uk" + git config user.name "sanger-tol-bot" git config push.default upstream git add . git status From 2aa9f869d0980fbaad83e8e2e7fcaeb3bce39fda Mon Sep 17 00:00:00 2001 From: Matthieu Muffato Date: Fri, 18 Aug 2023 16:09:54 +0100 Subject: [PATCH 014/123] New tolsoft details (#57) * Use the same name as our GitHub user to allow linking * Use the same email as our GitHub user to allow linking * Renamed the secret for consistency * Allow running the action on this branch * Messed up with the indentation * fixup! Allow running the action on this branch * Hardcode the issue number to allow checking out the code * Wrong issue number * [automated] Fix linting with Prettier * Restored the original condition * Fixed wrong user name Co-authored-by: Guoying Qi <729395+gq1@users.noreply.github.com> --------- Co-authored-by: sanger-tolsoft <105875386+sanger-tolsoft@users.noreply.github.com> Co-authored-by: Guoying Qi <729395+gq1@users.noreply.github.com> --- .github/workflows/fix-linting.yml | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml index dfb9b87..010b9f1 100644 --- a/.github/workflows/fix-linting.yml +++ b/.github/workflows/fix-linting.yml @@ -8,21 +8,21 @@ jobs: # Only run if comment is on a PR with the main repo, and if it contains the magic keywords if: > contains(github.event.comment.html_url, '/pull/') && - contains(github.event.comment.body, '@sanger-tol-bot fix linting') && + contains(github.event.comment.body, '@sanger-tolsoft fix linting') && github.repository == 'sanger-tol/variantcalling' runs-on: ubuntu-latest steps: - # Use the @sanger-tol-bot token to check out so we can push later + # Use the @sanger-tolsoft token to check out so we can push later - uses: actions/checkout@v3 with: - token: ${{ secrets.sangertol_bot_access_token }} + token: ${{ secrets.sangertolsoft_access_token }} # Action runs on the issue comment, so we don't get the PR by default # Use the gh cli to check out the PR - name: Checkout Pull Request run: gh pr checkout ${{ github.event.issue.number }} env: - GITHUB_TOKEN: ${{ secrets.sangertol_bot_access_token }} + GITHUB_TOKEN: ${{ secrets.sangertolsoft_access_token }} - uses: actions/setup-node@v3 @@ -46,8 +46,8 @@ jobs: - name: Commit & push changes if: steps.prettier_status.outputs.result == 'fail' run: | - git config user.email "tolsoft@sanger.ac.uk" - git config user.name "sanger-tol-bot" + git config user.email "105875386+sanger-tolsoft@users.noreply.github.com" + git config user.name "sanger-tolsoft" git config push.default upstream git add . git status From febbcff77ed35113dac3eeb0615460559d0565b1 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 11:31:07 +0100 Subject: [PATCH 015/123] nf-core lint --fix files_unchanged ( nf-core 2.8) --- .github/workflows/linting.yml | 2 +- lib/NfcoreSchema.groovy | 4 +++- 2 files changed, 4 insertions(+), 2 deletions(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 858d622..888cb4b 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -78,7 +78,7 @@ jobs: - uses: actions/setup-python@v4 with: - python-version: "3.7" + python-version: "3.8" architecture: "x64" - name: Install dependencies diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy index 33cd4f6..9b34804 100755 --- a/lib/NfcoreSchema.groovy +++ b/lib/NfcoreSchema.groovy @@ -2,6 +2,7 @@ // This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template. // +import nextflow.Nextflow import org.everit.json.schema.Schema import org.everit.json.schema.loader.SchemaLoader import org.everit.json.schema.ValidationException @@ -83,6 +84,7 @@ class NfcoreSchema { 'stub-run', 'test', 'w', + 'with-apptainer', 'with-charliecloud', 'with-conda', 'with-dag', @@ -177,7 +179,7 @@ class NfcoreSchema { } if (has_error) { - System.exit(1) + Nextflow.error('Exiting!') } } From 5fc66d604e6affd030878532ae8818357ddf6419 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 11:33:28 +0100 Subject: [PATCH 016/123] nf-core modules update bcftools/concat --- modules.json | 2 +- modules/nf-core/bcftools/concat/main.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/modules.json b/modules.json index 5ab5a99..e3332ee 100644 --- a/modules.json +++ b/modules.json @@ -7,7 +7,7 @@ "nf-core": { "bcftools/concat": { "branch": "master", - "git_sha": "fa12afdf5874c1d11e4a20efe81c97935e8eea24", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] }, "cat/cat": { diff --git a/modules/nf-core/bcftools/concat/main.nf b/modules/nf-core/bcftools/concat/main.nf index de9ba67..244a42c 100644 --- a/modules/nf-core/bcftools/concat/main.nf +++ b/modules/nf-core/bcftools/concat/main.nf @@ -5,7 +5,7 @@ process BCFTOOLS_CONCAT { conda "bioconda::bcftools=1.17" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/bcftools:1.17--haef29d1_0': - 'quay.io/biocontainers/bcftools:1.17--haef29d1_0' }" + 'biocontainers/bcftools:1.17--haef29d1_0' }" input: tuple val(meta), path(vcfs), path(tbi) From 3c7c11ae1a65fa344658dd560f85e78abd097b39 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 11:34:23 +0100 Subject: [PATCH 017/123] nf-core modules update cat/cat --- modules.json | 2 +- modules/nf-core/cat/cat/main.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/modules.json b/modules.json index e3332ee..7aa08b7 100644 --- a/modules.json +++ b/modules.json @@ -12,7 +12,7 @@ }, "cat/cat": { "branch": "master", - "git_sha": "0f8a77ff00e65eaeebc509b8156eaa983192474b", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] }, "custom/dumpsoftwareversions": { diff --git a/modules/nf-core/cat/cat/main.nf b/modules/nf-core/cat/cat/main.nf index 840af4b..9f06221 100644 --- a/modules/nf-core/cat/cat/main.nf +++ b/modules/nf-core/cat/cat/main.nf @@ -5,7 +5,7 @@ process CAT_CAT { conda "conda-forge::pigz=2.3.4" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/pigz:2.3.4' : - 'quay.io/biocontainers/pigz:2.3.4' }" + 'biocontainers/pigz:2.3.4' }" input: tuple val(meta), path(files_in) From bed22982c36481de35e7f52dd58f545df661e1c2 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 11:36:39 +0100 Subject: [PATCH 018/123] nf-core modules update custom/dumpsoftwareversions --- modules.json | 2 +- modules/nf-core/custom/dumpsoftwareversions/main.nf | 6 +++--- modules/nf-core/custom/dumpsoftwareversions/meta.yml | 2 ++ .../dumpsoftwareversions/templates/dumpsoftwareversions.py | 0 4 files changed, 6 insertions(+), 4 deletions(-) mode change 100644 => 100755 modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py diff --git a/modules.json b/modules.json index 7aa08b7..24c99bd 100644 --- a/modules.json +++ b/modules.json @@ -17,7 +17,7 @@ }, "custom/dumpsoftwareversions": { "branch": "master", - "git_sha": "7101db4432d3268b7fcb5b8f75fa0a022dc5561b", + "git_sha": "05c280924b6c768d484c7c443dad5e605c4ff4b4", "installed_by": ["modules"] }, "deepvariant": { diff --git a/modules/nf-core/custom/dumpsoftwareversions/main.nf b/modules/nf-core/custom/dumpsoftwareversions/main.nf index 800a609..c9d014b 100644 --- a/modules/nf-core/custom/dumpsoftwareversions/main.nf +++ b/modules/nf-core/custom/dumpsoftwareversions/main.nf @@ -2,10 +2,10 @@ process CUSTOM_DUMPSOFTWAREVERSIONS { label 'process_single' // Requires `pyyaml` which does not have a dedicated container but is in the MultiQC container - conda "bioconda::multiqc=1.14" + conda "bioconda::multiqc=1.15" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/multiqc:1.14--pyhdfd78af_0' : - 'quay.io/biocontainers/multiqc:1.14--pyhdfd78af_0' }" + 'https://depot.galaxyproject.org/singularity/multiqc:1.15--pyhdfd78af_0' : + 'biocontainers/multiqc:1.15--pyhdfd78af_0' }" input: path versions diff --git a/modules/nf-core/custom/dumpsoftwareversions/meta.yml b/modules/nf-core/custom/dumpsoftwareversions/meta.yml index 60b546a..c32657d 100644 --- a/modules/nf-core/custom/dumpsoftwareversions/meta.yml +++ b/modules/nf-core/custom/dumpsoftwareversions/meta.yml @@ -1,7 +1,9 @@ +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/yaml-schema.json name: custom_dumpsoftwareversions description: Custom module used to dump software versions within the nf-core pipeline template keywords: - custom + - dump - version tools: - custom: diff --git a/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py old mode 100644 new mode 100755 From c9b4e23fc2abbc4f905a135b18f0e1d47c5609b8 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 13:38:15 +0100 Subject: [PATCH 019/123] nf-core modules update deepvariant --- modules.json | 5 ++-- modules/nf-core/deepvariant/main.nf | 36 +++++++++++++++++----------- modules/nf-core/deepvariant/meta.yml | 17 +++++++++++++ 3 files changed, 41 insertions(+), 17 deletions(-) diff --git a/modules.json b/modules.json index 24c99bd..297798d 100644 --- a/modules.json +++ b/modules.json @@ -22,9 +22,8 @@ }, "deepvariant": { "branch": "master", - "git_sha": "58b5e78506e66f7ecd610fa825890ed9fb98b793", - "installed_by": ["modules"], - "patch": "modules/nf-core/deepvariant/deepvariant.diff" + "git_sha": "ed67f2fadd6d2a155b296f728e6b1f8c92ddc1a6", + "installed_by": ["modules"] }, "samtools/faidx": { "branch": "master", diff --git a/modules/nf-core/deepvariant/main.nf b/modules/nf-core/deepvariant/main.nf index 434fcc0..2d5c480 100644 --- a/modules/nf-core/deepvariant/main.nf +++ b/modules/nf-core/deepvariant/main.nf @@ -1,32 +1,33 @@ process DEEPVARIANT { tag "$meta.id" - label 'process_medium' + label 'process_high' - container "google/deepvariant:1.4.0" - - // Exit if running this module with -profile conda / -profile mamba - if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { - exit 1, "DEEPVARIANT module does not support Conda. Please use Docker / Singularity / Podman instead." - } + container "nf-core/deepvariant:1.5.0" input: tuple val(meta), path(input), path(index), path(intervals) - path(fasta) - path(fai) - path(gzi) + tuple val(meta2), path(fasta) + tuple val(meta3), path(fai) + tuple val(meta4), path(gzi) output: - tuple val(meta), path("${prefix}.vcf.gz") , emit: vcf - tuple val(meta), path("${prefix}.g.vcf.gz"), emit: gvcf - path "versions.yml" , emit: versions + tuple val(meta), path("${prefix}.vcf.gz") , emit: vcf + tuple val(meta), path("${prefix}.vcf.gz.tbi") , emit: vcf_tbi + tuple val(meta), path("${prefix}.g.vcf.gz") , emit: gvcf + tuple val(meta), path("${prefix}.g.vcf.gz.tbi"), emit: gvcf_tbi + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when script: + // Exit if running this module with -profile conda / -profile mamba + if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { + error "DEEPVARIANT module does not support Conda. Please use Docker / Singularity / Podman instead." + } def args = task.ext.args ?: '' prefix = task.ext.prefix ?: "${meta.id}" - def regions = intervals ? "--regions ${intervals}" : "" + def regions = intervals ? "--regions=${intervals}" : "" """ /opt/deepvariant/bin/run_deepvariant \\ @@ -36,6 +37,7 @@ process DEEPVARIANT { --output_gvcf=${prefix}.g.vcf.gz \\ ${args} \\ ${regions} \\ + --intermediate_results_dir=. \\ --num_shards=${task.cpus} cat <<-END_VERSIONS > versions.yml @@ -45,10 +47,16 @@ process DEEPVARIANT { """ stub: + // Exit if running this module with -profile conda / -profile mamba + if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { + error "DEEPVARIANT module does not support Conda. Please use Docker / Singularity / Podman instead." + } prefix = task.ext.prefix ?: "${meta.id}" """ touch ${prefix}.vcf.gz + touch ${prefix}.vcf.gz.tbi touch ${prefix}.g.vcf.gz + touch ${prefix}.g.vcf.gz.tbi cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/nf-core/deepvariant/meta.yml b/modules/nf-core/deepvariant/meta.yml index 63868b2..c7d11ae 100644 --- a/modules/nf-core/deepvariant/meta.yml +++ b/modules/nf-core/deepvariant/meta.yml @@ -3,6 +3,7 @@ description: DeepVariant is an analysis pipeline that uses a deep neural network keywords: - variant calling - machine learning + - neural network tools: - deepvariant: description: DeepVariant is an analysis pipeline that uses a deep neural network to call genetic variants from next-generation DNA sequencing data @@ -30,14 +31,29 @@ input: type: file description: Interval file for targeted regions pattern: "*.bed" + - meta2: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'genome' ] - fasta: type: file description: The reference fasta file pattern: "*.fasta" + - meta3: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'genome' ] - fai: type: file description: Index of reference fasta file pattern: "*.fai" + - meta4: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'genome' ] - gzi: type: file description: GZI index of reference fasta file @@ -64,3 +80,4 @@ output: authors: - "@abhi18av" + - "@ramprasadn" From 025297e1faf42bc1229ba2f7ae198cc274f96a98 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 19:20:25 +0100 Subject: [PATCH 020/123] Add pipeline template yml file --- pipeline_template.yml | 3 +++ 1 file changed, 3 insertions(+) create mode 100644 pipeline_template.yml diff --git a/pipeline_template.yml b/pipeline_template.yml new file mode 100644 index 0000000..0aa7398 --- /dev/null +++ b/pipeline_template.yml @@ -0,0 +1,3 @@ +prefix: sanger-tol +skip: + - igenomes From 77890858729c07eb0827c2a7d9c0431e67082790 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Sep 2023 19:23:52 +0100 Subject: [PATCH 021/123] add pipeline template file --- pipeline_template.yml | 3 +++ 1 file changed, 3 insertions(+) create mode 100644 pipeline_template.yml diff --git a/pipeline_template.yml b/pipeline_template.yml new file mode 100644 index 0000000..0aa7398 --- /dev/null +++ b/pipeline_template.yml @@ -0,0 +1,3 @@ +prefix: sanger-tol +skip: + - igenomes From 7a3aeb87f70f6450555a46dc8ba45fb03488a014 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 15:10:24 +0100 Subject: [PATCH 022/123] Template update for nf-core/tools version 2.8 --- .editorconfig | 2 +- .github/CONTRIBUTING.md | 6 +- .github/ISSUE_TEMPLATE/bug_report.yml | 3 +- .github/PULL_REQUEST_TEMPLATE.md | 4 +- .github/workflows/branch.yml | 18 +-- .github/workflows/clean-up.yml | 24 ++++ .github/workflows/linting.yml | 2 +- .github/workflows/sangerfulltest.yml | 37 ----- .github/workflows/sangertest.yml | 24 ---- .nf-core.yml | 11 +- .pre-commit-config.yaml | 5 + CHANGELOG.md | 2 +- CITATIONS.md | 5 + LICENSE | 2 +- README.md | 65 +++++---- assets/email_template.html | 2 +- assets/multiqc_config.yml | 12 ++ assets/schema_input.json | 2 +- assets/sendmail_template.txt | 25 ++++ bin/check_samplesheet.py | 3 - conf/base.config | 2 +- conf/modules.config | 4 + conf/test_full.config | 2 + docs/images/mqc_fastqc_adapter.png | Bin 0 -> 23458 bytes docs/images/mqc_fastqc_counts.png | Bin 0 -> 33918 bytes docs/images/mqc_fastqc_quality.png | Bin 0 -> 55769 bytes docs/output.md | 41 +++++- docs/usage.md | 132 ++++++------------ lib/NfcoreSchema.groovy | 4 +- lib/NfcoreTemplate.groovy | 38 ++++- lib/WorkflowMain.groovy | 18 ++- lib/WorkflowVariantcalling.groovy | 49 ++++++- main.nf | 3 +- modules.json | 12 +- modules/local/samplesheet_check.nf | 2 +- .../custom/dumpsoftwareversions/meta.yml | 2 + .../templates/dumpsoftwareversions.py | 3 +- modules/nf-core/fastqc/main.nf | 51 +++++++ modules/nf-core/fastqc/meta.yml | 52 +++++++ modules/nf-core/multiqc/main.nf | 53 +++++++ modules/nf-core/multiqc/meta.yml | 56 ++++++++ nextflow.config | 44 +++++- nextflow_schema.json | 34 ++++- pipeline_template.yml | 2 +- tower.yml | 5 + workflows/variantcalling.nf | 42 +++++- 46 files changed, 663 insertions(+), 242 deletions(-) create mode 100644 .github/workflows/clean-up.yml delete mode 100644 .github/workflows/sangerfulltest.yml delete mode 100644 .github/workflows/sangertest.yml create mode 100644 .pre-commit-config.yaml create mode 100644 assets/multiqc_config.yml create mode 100755 docs/images/mqc_fastqc_adapter.png create mode 100755 docs/images/mqc_fastqc_counts.png create mode 100755 docs/images/mqc_fastqc_quality.png create mode 100644 modules/nf-core/fastqc/main.nf create mode 100644 modules/nf-core/fastqc/meta.yml create mode 100644 modules/nf-core/multiqc/main.nf create mode 100644 modules/nf-core/multiqc/meta.yml create mode 100644 tower.yml diff --git a/.editorconfig b/.editorconfig index b78de6e..b6b3190 100644 --- a/.editorconfig +++ b/.editorconfig @@ -8,7 +8,7 @@ trim_trailing_whitespace = true indent_size = 4 indent_style = space -[*.{md,yml,yaml,html,css,scss,js,cff}] +[*.{md,yml,yaml,html,css,scss,js}] indent_size = 2 # These files are edited and tested upstream in nf-core/modules diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 61c8548..fb9e100 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -46,9 +46,9 @@ These tests are run both with the latest available version of `Nextflow` and als :warning: Only in the unlikely and regretful event of a release happening with a bug. -- On your own fork, make a new branch `patch` based on `upstream/main`. +- On your own fork, make a new branch `patch` based on `upstream/master`. - Fix the bug, and bump version (X.Y.Z+1). -- A PR should be made on `main` from patch to directly this particular bug. +- A PR should be made on `master` from patch to directly this particular bug. ## Pipeline contribution conventions @@ -66,6 +66,8 @@ If you wish to contribute a new step, please use the following coding standards: 6. Add sanity checks and validation for all relevant parameters. 7. Perform local tests to validate that the new code works as expected. 8. If applicable, add a new test command in `.github/workflow/ci.yml`. +9. Update MultiQC config `assets/multiqc_config.yml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module. +10. Add a description of the output files and if relevant any appropriate images from the MultiQC report to `docs/output.md`. ### Default values diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml index fa96bcf..1df5965 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.yml +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -44,7 +44,8 @@ body: * Executor _(eg. slurm, local, awsbatch)_ - * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter or Charliecloud)_ + * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter, Charliecloud, + or Apptainer)_ * OS _(eg. CentOS Linux, macOS, Linux Mint)_ diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 23d7977..d8affa1 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -8,14 +8,14 @@ These are the most common things requested on pull requests (PRs). Remember that PRs should be made against the dev branch, unless you're preparing a pipeline release. -Learn more about contributing: [CONTRIBUTING.md](https://github.com/sanger-tol/variantcalling/tree/main/.github/CONTRIBUTING.md) +Learn more about contributing: [CONTRIBUTING.md](https://github.com/sanger-tol/variantcalling/tree/master/.github/CONTRIBUTING.md) --> ## PR checklist - [ ] This comment contains a description of changes (with reason). - [ ] If you've fixed a bug or added code that should be tested, add tests! -- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/sanger-tol/variantcalling/tree/main/.github/CONTRIBUTING.md) +- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/sanger-tol/variantcalling/tree/master/.github/CONTRIBUTING.md) - [ ] Make sure your code lints (`nf-core lint`). - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). - [ ] Usage Documentation in `docs/usage.md` is updated. diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index bd3eccf..5defeb5 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -1,19 +1,19 @@ name: nf-core branch protection -# This workflow is triggered on PRs to main branch on the repository -# It fails when someone tries to make a PR against the nf-core `main` branch instead of `dev` +# This workflow is triggered on PRs to master branch on the repository +# It fails when someone tries to make a PR against the nf-core `master` branch instead of `dev` on: pull_request_target: - branches: [main] + branches: [master] jobs: test: runs-on: ubuntu-latest steps: - # PRs to the nf-core repo main branch are only ok if coming from the nf-core repo `dev` or any `patch` branches + # PRs to the nf-core repo master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches - name: Check PRs if: github.repository == 'sanger-tol/variantcalling' run: | - { [[ ${{github.event.pull_request.head.repo.full_name }} == sanger-tol/variantcalling ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + { [[ ${{github.event.pull_request.head.repo.full_name }} == sanger-tol/variantcalling ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] # If the above check failed, post a comment on the PR explaining the failure # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets @@ -22,7 +22,7 @@ jobs: uses: mshick/add-pr-comment@v1 with: message: | - ## This PR is against the `main` branch :x: + ## This PR is against the `master` branch :x: * Do not close this PR * Click _Edit_ and change the `base` to `dev` @@ -32,9 +32,9 @@ jobs: Hi @${{ github.event.pull_request.user.login }}, - It looks like this pull-request is has been made against the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `main` branch. - The `main` branch on nf-core repositories should always contain code from the latest release. - Because of this, PRs to `main` are only allowed if they come from the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `dev` branch. + It looks like this pull-request is has been made against the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `master` branch. + The `master` branch on nf-core repositories should always contain code from the latest release. + Because of this, PRs to `master` are only allowed if they come from the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `dev` branch. You do not need to close this PR, you can change the target branch to `dev` by clicking the _"Edit"_ button at the top of this page. Note that even after this, the test will continue to show as failing until you push a new commit. diff --git a/.github/workflows/clean-up.yml b/.github/workflows/clean-up.yml new file mode 100644 index 0000000..694e90e --- /dev/null +++ b/.github/workflows/clean-up.yml @@ -0,0 +1,24 @@ +name: "Close user-tagged issues and PRs" +on: + schedule: + - cron: "0 0 * * 0" # Once a week + +jobs: + clean-up: + runs-on: ubuntu-latest + permissions: + issues: write + pull-requests: write + steps: + - uses: actions/stale@v7 + with: + stale-issue-message: "This issue has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor. Remove stale label or add a comment otherwise this issue will be closed in 20 days." + stale-pr-message: "This PR has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor. Remove stale label or add a comment if it is still useful." + close-issue-message: "This issue was closed because it has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor and then staled for 20 days with no activity." + days-before-stale: 30 + days-before-close: 20 + days-before-pr-close: -1 + any-of-labels: "awaiting-changes,awaiting-feedback" + exempt-issue-labels: "WIP" + exempt-pr-labels: "WIP" + repo-token: "${{ secrets.GITHUB_TOKEN }}" diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 858d622..888cb4b 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -78,7 +78,7 @@ jobs: - uses: actions/setup-python@v4 with: - python-version: "3.7" + python-version: "3.8" architecture: "x64" - name: Install dependencies diff --git a/.github/workflows/sangerfulltest.yml b/.github/workflows/sangerfulltest.yml deleted file mode 100644 index 9d266be..0000000 --- a/.github/workflows/sangerfulltest.yml +++ /dev/null @@ -1,37 +0,0 @@ -name: sanger-tol LSF full size tests - -on: - push: - branches: - - main - - dev - workflow_dispatch: -jobs: - run-tower: - name: Run LSF full size tests - runs-on: ubuntu-latest - steps: - - name: Sets env vars for push - run: | - echo "REVISION=${{ github.sha }}" >> $GITHUB_ENV - if: github.event_name == 'push' - - - name: Sets env vars for workflow_dispatch - run: | - echo "REVISION=${{ github.sha }}" >> $GITHUB_ENV - if: github.event_name == 'workflow_dispatch' - - - name: Launch workflow via tower - uses: nf-core/tower-action@v2 - with: - workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} - access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} - compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - pipeline: ${{ github.repository }} - revision: ${{ env.REVISION }} - workdir: ${{ secrets.TOWER_WORKDIR_PARENT }}/work/${{ github.repository }}/work-${{ env.REVISION }} - parameters: | - { - "outdir": "${{ secrets.TOWER_WORKDIR_PARENT }}/results/${{ github.repository }}/results-${{ env.REVISION }}", - } - profiles: test_full,sanger,singularity diff --git a/.github/workflows/sangertest.yml b/.github/workflows/sangertest.yml deleted file mode 100644 index f1395fa..0000000 --- a/.github/workflows/sangertest.yml +++ /dev/null @@ -1,24 +0,0 @@ -name: sanger-tol LSF tests - -on: - workflow_dispatch: - -jobs: - run-tower: - name: Run LSF tests - runs-on: ubuntu-latest - steps: - - name: Launch workflow via tower - uses: nf-core/tower-action@v2 - with: - workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} - access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} - compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - pipeline: ${{ github.repository }} - revision: ${{ github.sha }} - workdir: ${{ secrets.TOWER_WORKDIR_PARENT }}/work/${{ github.repository }}/work-${{ github.sha }} - parameters: | - { - "outdir": "${{ secrets.TOWER_WORKDIR_PARENT }}/results/${{ github.repository }}/results-${{ github.sha }}", - } - profiles: test,sanger,singularity diff --git a/.nf-core.yml b/.nf-core.yml index 9caa7b0..3de0af8 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -8,15 +8,8 @@ lint: - .github/ISSUE_TEMPLATE/config.yml - .github/workflows/awstest.yml - .github/workflows/awsfulltest.yml - - assets/multiqc_config.yml - - conf/igenomes.config - files_unchanged: - - .github/CONTRIBUTING.md - - assets/sendmail_template.txt - - lib/NfcoreTemplate.groovy - - .github/PULL_REQUEST_TEMPLATE.md - - .github/workflows/branch.yml - nextflow_config: - manifest.name - manifest.homePage + multiqc_config: + - report_comment diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml new file mode 100644 index 0000000..0c31cdb --- /dev/null +++ b/.pre-commit-config.yaml @@ -0,0 +1,5 @@ +repos: + - repo: https://github.com/pre-commit/mirrors-prettier + rev: "v2.7.1" + hooks: + - id: prettier diff --git a/CHANGELOG.md b/CHANGELOG.md index c290a15..84a9bdc 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## v1.0dev - [date] +## v1.0.0 - [date] Initial release of sanger-tol/variantcalling, created with the [nf-core](https://nf-co.re/) template. diff --git a/CITATIONS.md b/CITATIONS.md index 4abdbdc..0d722f8 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -10,6 +10,11 @@ ## Pipeline tools +- [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) + +- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) + > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. + ## Software packaging/containerisation tools - [Anaconda](https://anaconda.com) diff --git a/LICENSE b/LICENSE index 7e408f7..f5f150c 100644 --- a/LICENSE +++ b/LICENSE @@ -1,6 +1,6 @@ MIT License -Copyright (c) Guoying Qi +Copyright (c) @muffato, @gq1, @priyanka-surana Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal diff --git a/README.md b/README.md index fc0cdd2..6efa459 100644 --- a/README.md +++ b/README.md @@ -8,50 +8,63 @@ ## Introduction - +**sanger-tol/variantcalling** is a bioinformatics pipeline that ... -**sanger-tol/variantcalling** is a bioinformatics best-practice analysis pipeline for variant calling. + -The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It uses Docker/Singularity containers making installation trivial and results highly reproducible. The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. Where possible, these processes have been submitted to and installed from [nf-core/modules](https://github.com/nf-core/modules) in order to make them available to all nf-core pipelines, and to everyone within the Nextflow community! - - + + -On release, automated continuous integration tests run the pipeline on a full-sized dataset on the LSF infrastructure. This ensures that the pipeline runs on LSF, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources. +1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)) +2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/)) -## Pipeline summary +## Usage - +> **Note** +> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how +> to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) +> with `-profile test` before running the workflow on actual data. -## Quick Start + - > - The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`. - > - Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. - > - If you are using `singularity`, please use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. - > - If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs. +Now, you can run the pipeline using: -4. Start running your own analysis! + - +```bash +nextflow run sanger-tol/variantcalling \ + -profile \ + --input samplesheet.csv \ + --outdir +``` - ```bash - nextflow run sanger-tol/variantcalling --input samplesheet.csv --outdir --genome GRCh37 -profile - ``` +> **Warning:** +> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those +> provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; +> see [docs](https://nf-co.re/usage/configuration#custom-configuration-files). ## Credits -sanger-tol/variantcalling was originally written by Guoying Qi. +sanger-tol/variantcalling was originally written by @muffato, @gq1, @priyanka-surana. We thank the following people for their extensive assistance in the development of this pipeline: diff --git a/assets/email_template.html b/assets/email_template.html index 68aa3d4..7bf2fb1 100644 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -4,7 +4,7 @@ - + sanger-tol/variantcalling Pipeline Report diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml new file mode 100644 index 0000000..34e2778 --- /dev/null +++ b/assets/multiqc_config.yml @@ -0,0 +1,12 @@ +report_comment: > + This report has been generated by the sanger-tol/variantcalling + analysis pipeline. +report_section_order: + "sanger-tol-variantcalling-methods-description": + order: -1000 + software_versions: + order: -1001 + "sanger-tol-variantcalling-summary": + order: -1002 + +export_plots: true diff --git a/assets/schema_input.json b/assets/schema_input.json index 50f9182..f97020b 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -1,6 +1,6 @@ { "$schema": "http://json-schema.org/draft-07/schema", - "$id": "https://raw.githubusercontent.com/sanger-tol/variantcalling/main/assets/schema_input.json", + "$id": "https://raw.githubusercontent.com/sanger-tol/variantcalling/master/assets/schema_input.json", "title": "sanger-tol/variantcalling pipeline - params.input schema", "description": "Schema for the file provided with params.input", "type": "array", diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt index 4ba6479..5e43653 100644 --- a/assets/sendmail_template.txt +++ b/assets/sendmail_template.txt @@ -25,4 +25,29 @@ Content-Disposition: inline; filename="sanger-tol-variantcalling_logo_light.png" flatten(). join( '\n' ) %> +<% +if (mqcFile){ +def mqcFileObj = new File("$mqcFile") +if (mqcFileObj.length() < mqcMaxSize){ +out << """ +--nfcoremimeboundary +Content-Type: text/html; name=\"multiqc_report\" +Content-Transfer-Encoding: base64 +Content-ID: +Content-Disposition: attachment; filename=\"${mqcFileObj.getName()}\" + +${mqcFileObj. + bytes. + encodeBase64(). + toString(). + tokenize( '\n' )*. + toList()*. + collate( 76 )*. + collect { it.join() }. + flatten(). + join( '\n' )} +""" +}} +%> + --nfcoremimeboundary-- diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py index 11b1557..4a758fe 100755 --- a/bin/check_samplesheet.py +++ b/bin/check_samplesheet.py @@ -158,9 +158,6 @@ def sniff_format(handle): peek = read_head(handle) handle.seek(0) sniffer = csv.Sniffer() - if not sniffer.has_header(peek): - logger.critical("The given sample sheet does not appear to contain a header.") - sys.exit(1) dialect = sniffer.sniff(peek) return dialect diff --git a/conf/base.config b/conf/base.config index df5f785..f43592d 100644 --- a/conf/base.config +++ b/conf/base.config @@ -15,7 +15,7 @@ process { memory = { check_max( 6.GB * task.attempt, 'memory' ) } time = { check_max( 4.h * task.attempt, 'time' ) } - errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' } + errorStrategy = { task.exitStatus in ((130..145) + 104) ? 'retry' : 'finish' } maxRetries = 1 maxErrors = '-1' diff --git a/conf/modules.config b/conf/modules.config index 60f6467..da58a5d 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -26,6 +26,10 @@ process { ] } + withName: FASTQC { + ext.args = '--quiet' + } + withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/conf/test_full.config b/conf/test_full.config index ac21d74..9f207b2 100644 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -10,6 +10,8 @@ ---------------------------------------------------------------------------------------- */ +cleanup = true + params { config_profile_name = 'Full test profile' config_profile_description = 'Full test dataset to check pipeline function' diff --git a/docs/images/mqc_fastqc_adapter.png b/docs/images/mqc_fastqc_adapter.png new file mode 100755 index 0000000000000000000000000000000000000000..361d0e47acfb424dea1f326590d1eb2f6dfa26b5 GIT binary patch literal 23458 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zXwhtT%ei{n#FzEH|C;yZ>+$!u_x#*+`=L8{b9SH^9&27u3G_Gxqxe`L2UJtdxghk z&-wzDFvLvW{chK5u3{n6GSKKy!P&C6w^IFpbD0bcp^A{{2lcLh_DXj@ybtYvc^;(2 M)78&qol`;+0Fu7JivR!s literal 0 HcmV?d00001 diff --git a/docs/output.md b/docs/output.md index 07a9d89..3666250 100644 --- a/docs/output.md +++ b/docs/output.md @@ -2,7 +2,7 @@ ## Introduction -This document describes the output produced by the pipeline. +This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory. @@ -12,8 +12,47 @@ The directories listed below will be created in the results directory after the The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: +- [FastQC](#fastqc) - Raw read QC +- [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline - [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution +### FastQC + +
+Output files + +- `fastqc/` + - `*_fastqc.html`: FastQC report containing quality metrics. + - `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images. + +
+ +[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your sequenced reads. It provides information about the quality score distribution across your reads, per base sequence content (%A/T/G/C), adapter contamination and overrepresented sequences. For further reading and documentation see the [FastQC help pages](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/). + +![MultiQC - FastQC sequence counts plot](images/mqc_fastqc_counts.png) + +![MultiQC - FastQC mean quality scores plot](images/mqc_fastqc_quality.png) + +![MultiQC - FastQC adapter content plot](images/mqc_fastqc_adapter.png) + +> **NB:** The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They may contain adapter sequence and potentially regions with low quality. + +### MultiQC + +
+Output files + +- `multiqc/` + - `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser. + - `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline. + - `multiqc_plots/`: directory containing static images from the report in various formats. + +
+ +[MultiQC](http://multiqc.info) is a visualization tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in the report data directory. + +Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQC. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. 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af798af51ca5607029e95d41f95012230ffcce75 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Dec 2023 09:55:49 +0000 Subject: [PATCH 119/123] update usage. --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index 131c5b8..4fbed55 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -63,7 +63,7 @@ work # Directory containing the nextflow working files The pipeline will split the input fasta file into smaller files to run DeepVariant parallel. You can set the minimum split fasta file size from the command line. For example to set the minimum size as 10K using `--split_fasta_cutoff 10000`. -If the input BAM/CRAM files are not aligned, please add `--align` in your command. +If the input BAM/CRAM files are not aligned, please add `--align` in your command. Please don't use this flag if the input files are already aligned because the current workflow will not align the aligned PacBio reads. ### Updating the pipeline From ce19b42469d26fa43cf5c58d65cd72a0e7d2947f Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Dec 2023 09:57:10 +0000 Subject: [PATCH 120/123] Add a name for previous release --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 62bb55c..e31522d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -49,7 +49,7 @@ Note, since the pipeline is using Nextflow DSL2, each process will be run with i > **NB:** Dependency has been **updated** if both old and new version information is present.
    **NB:** Dependency has been **added** if just the new version information is present.
    **NB:** Dependency has been **removed** if version information isn't present. -## [[1.0.0](https://github.com/sanger-tol/variantcalling/releases/tag/1.0.0)] - [2023-05-03] +## [[1.0.0](https://github.com/sanger-tol/variantcalling/releases/tag/1.0.0)] - Xia Yu - [2023-05-03] Initial release of sanger-tol/variantcalling, created with the [nf-core](https://nf-co.re/) tools. From 64eba1f4f68b30240353dcbb1bf5bd7e642a4c2d Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Dec 2023 10:13:03 +0000 Subject: [PATCH 121/123] Add citation file --- CITATION.cff | 30 ++++++++++++++++++++++++++++++ 1 file changed, 30 insertions(+) create mode 100644 CITATION.cff diff --git a/CITATION.cff b/CITATION.cff new file mode 100644 index 0000000..5b0fb48 --- /dev/null +++ b/CITATION.cff @@ -0,0 +1,30 @@ +# This CITATION.cff file was generated with cffinit. +# Visit https://bit.ly/cffinit to generate yours today! + +cff-version: 1.2.0 +title: sanger-tol/variantcalling v1.1.0 +message: >- + If you use this software, please cite it using the + metadata from this file. +type: software +authors: + - given-names: Guoying + family-names: Qi + affiliation: Wellcome Sanger Institute + orcid: "https://orcid.org/0000-0003-1262-8973" + - given-names: Priyanka + family-names: Surana + affiliation: Wellcome Sanger Institute + orcid: "https://orcid.org/0000-0002-7167-0875" + - given-names: Matthieu + family-names: Muffato + affiliation: Wellcome Sanger Institute + orcid: "https://orcid.org/0000-0002-7860-3560" +identifiers: + - type: doi + value: 10.5281/zenodo.7890528 +repository-code: "https://github.com/sanger-tol/variantcalling" +license: MIT +commit: TODO +version: 1.1.0 +date-released: "2022-12-20" From c287ef7180ef1ff4709375ad40254957559aad06 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Dec 2023 10:21:52 +0000 Subject: [PATCH 122/123] ignore cff file for EC lint check --- .github/workflows/linting.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 3f27dab..916c43a 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -22,7 +22,7 @@ jobs: run: npm install -g editorconfig-checker - name: Run ECLint check - run: editorconfig-checker -exclude README.md $(find .* -type f | grep -v '.git\|.py\|.md\|json\|yml\|yaml\|html\|css\|work\|.nextflow\|build\|nf_core.egg-info\|log.txt\|Makefile') + run: editorconfig-checker -exclude README.md $(find .* -type f | grep -v '.git\|.py\|.cff\|.md\|json\|yml\|yaml\|html\|css\|work\|.nextflow\|build\|nf_core.egg-info\|log.txt\|Makefile') Prettier: runs-on: ubuntu-latest From d3058e7f0fa496f6f2a6f33fbcda8a8b329d918d Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 20 Dec 2023 10:38:57 +0000 Subject: [PATCH 123/123] typo --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index e31522d..bbd01fb 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,7 +10,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Updated the CI procedure to use "sanger-tol" rather than "nf-core" names. - Renamed Sanger related Github CI test workflows. - nf-core template was updated from 2.7 to 2.8. -- Removed BAM/CRAM index files from the Sample sheets. +- Removed BAM/CRAM index files from the sample sheets. - Made fasta index file optional from the inputs. - Imported PacBio readmapping sub-workflows from [sanger-tol/readmapping pipeline](https://github.com/sanger-tol/readmapping/). Therefore, the pipeline can run on unaligned BAM/CRAM samples now. - Use VCFtools to calculate per site nucleotide diversity.

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diff --git a/docs/usage.md b/docs/usage.md index d337109..ff01839 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -69,6 +69,29 @@ work # Directory containing the nextflow working files # Other nextflow hidden files, eg. history of pipeline runs and old logs. ``` +If you wish to repeatedly use the same parameters for multiple runs, rather than specifying each flag in the command, you can specify these in a params file. + +Pipeline settings can be provided in a `yaml` or `json` file via `-params-file `. + +> ⚠️ Do not use `-c ` to specify parameters as this will result in errors. Custom config files specified with `-c` must only be used for [tuning process resource specifications](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources), other infrastructural tweaks (such as output directories), or module arguments (args). +> The above pipeline run specified with a params file in yaml format: + +```bash +nextflow run sanger-tol/variantcalling -profile docker -params-file params.yaml +``` + +with `params.yaml` containing: + +```yaml +input: './samplesheet.csv' +outdir: './results/' +genome: 'GRCh37' +input: 'data' +<...> +``` + +You can also generate such `YAML`/`JSON` files via [nf-core/launch](https://nf-co.re/launch). + ### Updating the pipeline When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline: @@ -83,7 +106,11 @@ It is a good idea to specify a pipeline version when running the pipeline on you First, go to the [sanger-tol/variantcalling releases page](https://github.com/sanger-tol/variantcalling/releases) and find the latest pipeline version - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. Of course, you can switch to another version by changing the number after the `-r` flag. -This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. +This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. For example, at the bottom of the MultiQC reports. + +To further assist in reproducbility, you can use share and re-use [parameter files](#running-the-pipeline) to repeat pipeline runs with the same settings without having to write out a command with every single parameter. + +> 💡 If you wish to share such profile (such as upload as supplementary material for academic publications), make sure to NOT include cluster specific paths to files, nor institutional specific profiles. ## Core Nextflow arguments @@ -93,7 +120,7 @@ This version number will be logged in reports when you run the pipeline, so that Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. -Several generic profiles are bundled with the pipeline which instruct the pipeline to use software packaged using different methods (Docker, Singularity, Podman, Shifter, Charliecloud, Conda) - see below. +Several generic profiles are bundled with the pipeline which instruct the pipeline to use software packaged using different methods (Docker, Singularity, Podman, Shifter, Charliecloud, Apptainer, Conda) - see below. > We highly recommend the use of Docker or Singularity containers for full pipeline reproducibility, however when this is not possible, Conda is also supported. @@ -117,8 +144,10 @@ If `-profile` is not specified, the pipeline will run locally and expect all sof - A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/) - `charliecloud` - A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/) +- `apptainer` + - A generic configuration profile to be used with [Apptainer](https://apptainer.org/) - `conda` - - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter or Charliecloud. + - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter, Charliecloud, or Apptainer. ### `-resume` @@ -136,102 +165,19 @@ Specify the path to a specific config file (this is a core Nextflow command). Se Whilst the default requirements set within the pipeline will hopefully work for most people and with most input data, you may find that you want to customise the compute resources that the pipeline requests. Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with any of the error codes specified [here](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L18) it will automatically be resubmitted with higher requests (2 x original, then 3 x original). If it still fails after the third attempt then the pipeline execution is stopped. -For example, if the nf-core/rnaseq pipeline is failing after multiple re-submissions of the `STAR_ALIGN` process due to an exit code of `137` this would indicate that there is an out of memory issue: - -```console -[62/149eb0] NOTE: Process `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) -- Execution is retried (1) -Error executing process > 'NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)' - -Caused by: - Process `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN (WT_REP1)` terminated with an error exit status (137) - -Command executed: - STAR \ - --genomeDir star \ - --readFilesIn WT_REP1_trimmed.fq.gz \ - --runThreadN 2 \ - --outFileNamePrefix WT_REP1. \ - - -Command exit status: - 137 - -Command output: - (empty) - -Command error: - .command.sh: line 9: 30 Killed STAR --genomeDir star --readFilesIn WT_REP1_trimmed.fq.gz --runThreadN 2 --outFileNamePrefix WT_REP1. -Work dir: - /home/pipelinetest/work/9d/172ca5881234073e8d76f2a19c88fb - -Tip: you can replicate the issue by changing to the process work dir and entering the command `bash .command.run` -``` - -#### For beginners - -A first step to bypass this error, you could try to increase the amount of CPUs, memory, and time for the whole pipeline. Therefor you can try to increase the resource for the parameters `--max_cpus`, `--max_memory`, and `--max_time`. Based on the error above, you have to increase the amount of memory. Therefore you can go to the [parameter documentation of rnaseq](https://nf-co.re/rnaseq/3.9/parameters) and scroll down to the `show hidden parameter` button to get the default value for `--max_memory`. In this case 128GB, you than can try to run your pipeline again with `--max_memory 200GB -resume` to skip all process, that were already calculated. If you can not increase the resource of the complete pipeline, you can try to adapt the resource for a single process as mentioned below. - -#### Advanced option on process level - -To bypass this error you would need to find exactly which resources are set by the `STAR_ALIGN` process. The quickest way is to search for `process STAR_ALIGN` in the [nf-core/rnaseq Github repo](https://github.com/nf-core/rnaseq/search?q=process+STAR_ALIGN). -We have standardised the structure of Nextflow DSL2 pipelines such that all module files will be present in the `modules/` directory and so, based on the search results, the file we want is `modules/nf-core/star/align/main.nf`. -If you click on the link to that file you will notice that there is a `label` directive at the top of the module that is set to [`label process_high`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/modules/nf-core/software/star/align/main.nf#L9). -The [Nextflow `label`](https://www.nextflow.io/docs/latest/process.html#label) directive allows us to organise workflow processes in separate groups which can be referenced in a configuration file to select and configure subset of processes having similar computing requirements. -The default values for the `process_high` label are set in the pipeline's [`base.config`](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L33-L37) which in this case is defined as 72GB. -Providing you haven't set any other standard nf-core parameters to **cap** the [maximum resources](https://nf-co.re/usage/configuration#max-resources) used by the pipeline then we can try and bypass the `STAR_ALIGN` process failure by creating a custom config file that sets at least 72GB of memory, in this case increased to 100GB. -The custom config below can then be provided to the pipeline via the [`-c`](#-c) parameter as highlighted in previous sections. - -```nextflow -process { - withName: 'NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN' { - memory = 100.GB - } -} -``` - -> **NB:** We specify the full process name i.e. `NFCORE_RNASEQ:RNASEQ:ALIGN_STAR:STAR_ALIGN` in the config file because this takes priority over the short name (`STAR_ALIGN`) and allows existing configuration using the full process name to be correctly overridden. -> -> If you get a warning suggesting that the process selector isn't recognised check that the process name has been specified correctly. - -### Updating containers (advanced users) - -The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. If for some reason you need to use a different version of a particular tool with the pipeline then you just need to identify the `process` name and override the Nextflow `container` definition for that process using the `withName` declaration. For example, in the [nf-core/viralrecon](https://nf-co.re/viralrecon) pipeline a tool called [Pangolin](https://github.com/cov-lineages/pangolin) has been used during the COVID-19 pandemic to assign lineages to SARS-CoV-2 genome sequenced samples. Given that the lineage assignments change quite frequently it doesn't make sense to re-release the nf-core/viralrecon everytime a new version of Pangolin has been released. However, you can override the default container used by the pipeline by creating a custom config file and passing it as a command-line argument via `-c custom.config`. - -1. Check the default version used by the pipeline in the module file for [Pangolin](https://github.com/nf-core/viralrecon/blob/a85d5969f9025409e3618d6c280ef15ce417df65/modules/nf-core/software/pangolin/main.nf#L14-L19) -2. Find the latest version of the Biocontainer available on [Quay.io](https://quay.io/repository/biocontainers/pangolin?tag=latest&tab=tags) -3. Create the custom config accordingly: - - - For Docker: +To change the resource requests, please see the [max resources](https://nf-co.re/docs/usage/configuration#max-resources) and [tuning workflow resources](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources) section of the nf-core website. - ```nextflow - process { - withName: PANGOLIN { - container = 'quay.io/biocontainers/pangolin:3.0.5--pyhdfd78af_0' - } - } - ``` +### Custom Containers - - For Singularity: +In some cases you may wish to change which container or conda environment a step of the pipeline uses for a particular tool. By default nf-core pipelines use containers and software from the [biocontainers](https://biocontainers.pro/) or [bioconda](https://bioconda.github.io/) projects. However in some cases the pipeline specified version maybe out of date. - ```nextflow - process { - withName: PANGOLIN { - container = 'https://depot.galaxyproject.org/singularity/pangolin:3.0.5--pyhdfd78af_0' - } - } - ``` +To use a different container from the default container or conda environment specified in a pipeline, please see the [updating tool versions](https://nf-co.re/docs/usage/configuration#updating-tool-versions) section of the nf-core website. - - For Conda: +### Custom Tool Arguments - ```nextflow - process { - withName: PANGOLIN { - conda = 'bioconda::pangolin=3.0.5' - } - } - ``` +A pipeline might not always support every possible argument or option of a particular tool used in pipeline. Fortunately, nf-core pipelines provide some freedom to users to insert additional parameters that the pipeline does not include by default. -> **NB:** If you wish to periodically update individual tool-specific results (e.g. Pangolin) generated by the pipeline then you must ensure to keep the `work/` directory otherwise the `-resume` ability of the pipeline will be compromised and it will restart from scratch. +To learn how to provide additional arguments to a particular tool of the pipeline, please see the [customising tool arguments](https://nf-co.re/docs/usage/configuration#customising-tool-arguments) section of the nf-core website. ### nf-core/configs diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy index 33cd4f6..9b34804 100755 --- a/lib/NfcoreSchema.groovy +++ b/lib/NfcoreSchema.groovy @@ -2,6 +2,7 @@ // This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template. // +import nextflow.Nextflow import org.everit.json.schema.Schema import org.everit.json.schema.loader.SchemaLoader import org.everit.json.schema.ValidationException @@ -83,6 +84,7 @@ class NfcoreSchema { 'stub-run', 'test', 'w', + 'with-apptainer', 'with-charliecloud', 'with-conda', 'with-dag', @@ -177,7 +179,7 @@ class NfcoreSchema { } if (has_error) { - System.exit(1) + Nextflow.error('Exiting!') } } diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 6ddfe19..2023f45 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -6,6 +6,18 @@ import org.yaml.snakeyaml.Yaml class NfcoreTemplate { + // + // Check AWS Batch related parameters have been specified correctly + // + public static void awsBatch(workflow, params) { + if (workflow.profile.contains('awsbatch')) { + // Check params.awsqueue and params.awsregion have been set if running on AWSBatch + assert (params.awsqueue && params.awsregion) : "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" + // Check outdir paths to be S3 buckets if running on AWSBatch + assert params.outdir.startsWith('s3:') : "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" + } + } + // // Warn if a -profile or Nextflow config has not been provided to run the pipeline // @@ -42,7 +54,7 @@ class NfcoreTemplate { // // Construct and send completion email // - public static void email(workflow, params, summary_params, projectDir, log) { + public static void email(workflow, params, summary_params, projectDir, log, multiqc_report=[]) { // Set up the e-mail variables def subject = "[$workflow.manifest.name] Successful: $workflow.runName" @@ -80,6 +92,24 @@ class NfcoreTemplate { email_fields['projectDir'] = workflow.projectDir email_fields['summary'] = summary << misc_fields + // On success try attach the multiqc report + def mqc_report = null + try { + if (workflow.success) { + mqc_report = multiqc_report.getVal() + if (mqc_report.getClass() == ArrayList && mqc_report.size() >= 1) { + if (mqc_report.size() > 1) { + log.warn "[$workflow.manifest.name] Found multiple reports from process 'MULTIQC', will use only one" + } + mqc_report = mqc_report[0] + } + } + } catch (all) { + if (multiqc_report) { + log.warn "[$workflow.manifest.name] Could not attach MultiQC report to summary email" + } + } + // Check if we are only sending emails on failure def email_address = params.email if (!params.email && params.email_on_fail && !workflow.success) { @@ -98,7 +128,8 @@ class NfcoreTemplate { def email_html = html_template.toString() // Render the sendmail template - def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir" ] + def max_multiqc_email_size = params.max_multiqc_email_size as nextflow.util.MemoryUnit + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ] def sf = new File("$projectDir/assets/sendmail_template.txt") def sendmail_template = engine.createTemplate(sf).make(smail_fields) def sendmail_html = sendmail_template.toString() @@ -114,6 +145,9 @@ class NfcoreTemplate { } catch (all) { // Catch failures and try with plaintext def mail_cmd = [ 'mail', '-s', subject, '--content-type=text/html', email_address ] + if ( mqc_report.size() <= max_multiqc_email_size.toBytes() ) { + mail_cmd += [ '-A', mqc_report ] + } mail_cmd.execute() << email_html log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (mail)-" } diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index a25a7d5..56ba5cb 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -2,6 +2,8 @@ // This file holds several functions specific to the main.nf workflow in the sanger-tol/variantcalling pipeline // +import nextflow.Nextflow + class WorkflowMain { // @@ -15,13 +17,13 @@ class WorkflowMain { "* The nf-core framework\n" + " https://doi.org/10.1038/s41587-020-0439-x\n\n" + "* Software dependencies\n" + - " https://github.com/${workflow.manifest.name}/blob/main/CITATIONS.md" + " https://github.com/${workflow.manifest.name}/blob/master/CITATIONS.md" } // // Generate help string // - public static String help(workflow, params, log) { + public static String help(workflow, params) { def command = "nextflow run ${workflow.manifest.name} --input samplesheet.csv --fasta reference.fa -profile docker" def help_string = '' help_string += NfcoreTemplate.logo(workflow, params.monochrome_logs) @@ -34,7 +36,7 @@ class WorkflowMain { // // Generate parameter summary log string // - public static String paramsSummaryLog(workflow, params, log) { + public static String paramsSummaryLog(workflow, params) { def summary_log = '' summary_log += NfcoreTemplate.logo(workflow, params.monochrome_logs) summary_log += NfcoreSchema.paramsSummaryLog(workflow, params) @@ -49,7 +51,7 @@ class WorkflowMain { public static void initialise(workflow, params, log) { // Print help to screen if required if (params.help) { - log.info help(workflow, params, log) + log.info help(workflow, params) System.exit(0) } @@ -61,7 +63,7 @@ class WorkflowMain { } // Print parameter summary log to screen - log.info paramsSummaryLog(workflow, params, log) + log.info paramsSummaryLog(workflow, params) // Validate workflow parameters via the JSON schema if (params.validate_params) { @@ -76,10 +78,12 @@ class WorkflowMain { Utils.checkCondaChannels(log) } + // Check AWS batch settings + NfcoreTemplate.awsBatch(workflow, params) + // Check input has been provided if (!params.input) { - log.error "Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.csv'" - System.exit(1) + Nextflow.error("Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.csv'") } } } diff --git a/lib/WorkflowVariantcalling.groovy b/lib/WorkflowVariantcalling.groovy index dad2c91..f64085e 100755 --- a/lib/WorkflowVariantcalling.groovy +++ b/lib/WorkflowVariantcalling.groovy @@ -2,6 +2,7 @@ // This file holds several functions specific to the workflow/variantcalling.nf in the sanger-tol/variantcalling pipeline // +import nextflow.Nextflow import groovy.text.SimpleTemplateEngine class WorkflowVariantcalling { @@ -10,11 +11,53 @@ class WorkflowVariantcalling { // Check and validate parameters // public static void initialise(params, log) { + if (!params.fasta) { - log.error "Genome fasta file not specified with e.g. '--fasta genome.fa' or via a detectable config file." - System.exit(1) + Nextflow.error "Genome fasta file not specified with e.g. '--fasta genome.fa' or via a detectable config file." } } -} + // + // Get workflow summary for MultiQC + // + public static String paramsSummaryMultiqc(workflow, summary) { + String summary_section = '' + for (group in summary.keySet()) { + def group_params = summary.get(group) // This gets the parameters of that particular group + if (group_params) { + summary_section += "

$group

\n" + summary_section += "
\n" + for (param in group_params.keySet()) { + summary_section += "
$param
${group_params.get(param) ?: 'N/A'}
\n" + } + summary_section += "
\n" + } + } + + String yaml_file_text = "id: '${workflow.manifest.name.replace('/','-')}-summary'\n" + yaml_file_text += "description: ' - this information is collected when the pipeline is started.'\n" + yaml_file_text += "section_name: '${workflow.manifest.name} Workflow Summary'\n" + yaml_file_text += "section_href: 'https://github.com/${workflow.manifest.name}'\n" + yaml_file_text += "plot_type: 'html'\n" + yaml_file_text += "data: |\n" + yaml_file_text += "${summary_section}" + return yaml_file_text + } + + public static String methodsDescriptionText(run_workflow, mqc_methods_yaml) { + // Convert to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file + def meta = [:] + meta.workflow = run_workflow.toMap() + meta["manifest_map"] = run_workflow.manifest.toMap() + + meta["doi_text"] = meta.manifest_map.doi ? "(doi: ${meta.manifest_map.doi})" : "" + meta["nodoi_text"] = meta.manifest_map.doi ? "": "
  • If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used.
    • " + + def methods_text = mqc_methods_yaml.text + + def engine = new SimpleTemplateEngine() + def description_html = engine.createTemplate(methods_text).make(meta) + + return description_html + }} diff --git a/main.nf b/main.nf index 61d8c25..7582b32 100644 --- a/main.nf +++ b/main.nf @@ -3,8 +3,7 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ sanger-tol/variantcalling ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - Github : https://github.com/sanger-tol/variantcalling ----------------------------------------------------------------------------------------- + Github : https://github.com/sanger-tol/variantcalling---------------------------------------------------------------------------------------- */ nextflow.enable.dsl = 2 diff --git a/modules.json b/modules.json index 8953ab5..4b03a7d 100644 --- a/modules.json +++ b/modules.json @@ -7,7 +7,17 @@ "nf-core": { "custom/dumpsoftwareversions": { "branch": "master", - "git_sha": "b6d4d476aee074311c89d82a69c1921bd70c8180", + "git_sha": "76cc4938c1f6ea5c7d83fed1eeffc146787f9543", + "installed_by": ["modules"] + }, + "fastqc": { + "branch": "master", + "git_sha": "c8e35eb2055c099720a75538d1b8adb3fb5a464c", + "installed_by": ["modules"] + }, + "multiqc": { + "branch": "master", + "git_sha": "f2d63bd5b68925f98f572eed70993d205cc694b7", "installed_by": ["modules"] } } diff --git a/modules/local/samplesheet_check.nf b/modules/local/samplesheet_check.nf index 264bee0..0505378 100644 --- a/modules/local/samplesheet_check.nf +++ b/modules/local/samplesheet_check.nf @@ -5,7 +5,7 @@ process SAMPLESHEET_CHECK { conda "conda-forge::python=3.8.3" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/python:3.8.3' : - 'quay.io/biocontainers/python:3.8.3' }" + 'biocontainers/python:3.8.3' }" input: path samplesheet diff --git a/modules/nf-core/custom/dumpsoftwareversions/meta.yml b/modules/nf-core/custom/dumpsoftwareversions/meta.yml index 60b546a..c32657d 100644 --- a/modules/nf-core/custom/dumpsoftwareversions/meta.yml +++ b/modules/nf-core/custom/dumpsoftwareversions/meta.yml @@ -1,7 +1,9 @@ +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/yaml-schema.json name: custom_dumpsoftwareversions description: Custom module used to dump software versions within the nf-core pipeline template keywords: - custom + - dump - version tools: - custom: diff --git a/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py index da03340..e55b8d4 100755 --- a/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py +++ b/modules/nf-core/custom/dumpsoftwareversions/templates/dumpsoftwareversions.py @@ -4,10 +4,11 @@ """Provide functions to merge multiple versions.yml files.""" -import yaml import platform from textwrap import dedent +import yaml + def _make_versions_html(versions): """Generate a tabular HTML output of all versions for MultiQC.""" diff --git a/modules/nf-core/fastqc/main.nf b/modules/nf-core/fastqc/main.nf new file mode 100644 index 0000000..9ae5838 --- /dev/null +++ b/modules/nf-core/fastqc/main.nf @@ -0,0 +1,51 @@ +process FASTQC { + tag "$meta.id" + label 'process_medium' + + conda "bioconda::fastqc=0.11.9" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/fastqc:0.11.9--0' : + 'quay.io/biocontainers/fastqc:0.11.9--0' }" + + input: + tuple val(meta), path(reads) + + output: + tuple val(meta), path("*.html"), emit: html + tuple val(meta), path("*.zip") , emit: zip + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // Make list of old name and new name pairs to use for renaming in the bash while loop + def old_new_pairs = reads instanceof Path || reads.size() == 1 ? [[ reads, "${prefix}.${reads.extension}" ]] : reads.withIndex().collect { entry, index -> [ entry, "${prefix}_${index + 1}.${entry.extension}" ] } + def rename_to = old_new_pairs*.join(' ').join(' ') + def renamed_files = old_new_pairs.collect{ old_name, new_name -> new_name }.join(' ') + """ + printf "%s %s\\n" $rename_to | while read old_name new_name; do + [ -f "\${new_name}" ] || ln -s \$old_name \$new_name + done + fastqc $args --threads $task.cpus $renamed_files + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" ) + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch ${prefix}.html + touch ${prefix}.zip + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + fastqc: \$( fastqc --version | sed -e "s/FastQC v//g" ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/fastqc/meta.yml b/modules/nf-core/fastqc/meta.yml new file mode 100644 index 0000000..4da5bb5 --- /dev/null +++ b/modules/nf-core/fastqc/meta.yml @@ -0,0 +1,52 @@ +name: fastqc +description: Run FastQC on sequenced reads +keywords: + - quality control + - qc + - adapters + - fastq +tools: + - fastqc: + description: | + FastQC gives general quality metrics about your reads. + It provides information about the quality score distribution + across your reads, the per base sequence content (%A/C/G/T). + You get information about adapter contamination and other + overrepresented sequences. + homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ + documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ + licence: ["GPL-2.0-only"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - html: + type: file + description: FastQC report + pattern: "*_{fastqc.html}" + - zip: + type: file + description: FastQC report archive + pattern: "*_{fastqc.zip}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@grst" + - "@ewels" + - "@FelixKrueger" diff --git a/modules/nf-core/multiqc/main.nf b/modules/nf-core/multiqc/main.nf new file mode 100644 index 0000000..4b60474 --- /dev/null +++ b/modules/nf-core/multiqc/main.nf @@ -0,0 +1,53 @@ +process MULTIQC { + label 'process_single' + + conda "bioconda::multiqc=1.14" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.14--pyhdfd78af_0' : + 'quay.io/biocontainers/multiqc:1.14--pyhdfd78af_0' }" + + input: + path multiqc_files, stageAs: "?/*" + path(multiqc_config) + path(extra_multiqc_config) + path(multiqc_logo) + + output: + path "*multiqc_report.html", emit: report + path "*_data" , emit: data + path "*_plots" , optional:true, emit: plots + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def config = multiqc_config ? "--config $multiqc_config" : '' + def extra_config = extra_multiqc_config ? "--config $extra_multiqc_config" : '' + """ + multiqc \\ + --force \\ + $args \\ + $config \\ + $extra_config \\ + . + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" ) + END_VERSIONS + """ + + stub: + """ + touch multiqc_data + touch multiqc_plots + touch multiqc_report.html + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/multiqc/meta.yml b/modules/nf-core/multiqc/meta.yml new file mode 100644 index 0000000..f93b5ee --- /dev/null +++ b/modules/nf-core/multiqc/meta.yml @@ -0,0 +1,56 @@ +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/yaml-schema.json +name: MultiQC +description: Aggregate results from bioinformatics analyses across many samples into a single report +keywords: + - QC + - bioinformatics tools + - Beautiful stand-alone HTML report +tools: + - multiqc: + description: | + MultiQC searches a given directory for analysis logs and compiles a HTML report. + It's a general use tool, perfect for summarising the output from numerous bioinformatics tools. + homepage: https://multiqc.info/ + documentation: https://multiqc.info/docs/ + licence: ["GPL-3.0-or-later"] + +input: + - multiqc_files: + type: file + description: | + List of reports / files recognised by MultiQC, for example the html and zip output of FastQC + - multiqc_config: + type: file + description: Optional config yml for MultiQC + pattern: "*.{yml,yaml}" + - extra_multiqc_config: + type: file + description: Second optional config yml for MultiQC. Will override common sections in multiqc_config. + pattern: "*.{yml,yaml}" + - multiqc_logo: + type: file + description: Optional logo file for MultiQC + pattern: "*.{png}" + +output: + - report: + type: file + description: MultiQC report file + pattern: "multiqc_report.html" + - data: + type: directory + description: MultiQC data dir + pattern: "multiqc_data" + - plots: + type: file + description: Plots created by MultiQC + pattern: "*_data" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@abhi18av" + - "@bunop" + - "@drpatelh" + - "@jfy133" diff --git a/nextflow.config b/nextflow.config index 0527f42..8f7d1d0 100644 --- a/nextflow.config +++ b/nextflow.config @@ -13,6 +13,13 @@ params { // Input options input = null +// MultiQC options + multiqc_config = null + multiqc_title = null + multiqc_logo = null + max_multiqc_email_size = '25.MB' + multiqc_methods_description = null + // Boilerplate options outdir = null tracedir = "${params.outdir}/pipeline_info" @@ -26,7 +33,7 @@ params { version = false validate_params = true show_hidden_params = false - schema_ignore_params = 'fasta' + schema_ignore_params = 'genomes' // Config options @@ -66,7 +73,11 @@ try { profiles { - debug { process.beforeScript = 'echo $HOSTNAME' } + debug { + dumpHashes = true + process.beforeScript = 'echo $HOSTNAME' + cleanup = false + } conda { conda.enabled = true docker.enabled = false @@ -74,6 +85,7 @@ profiles { podman.enabled = false shifter.enabled = false charliecloud.enabled = false + apptainer.enabled = false } mamba { conda.enabled = true @@ -83,14 +95,18 @@ profiles { podman.enabled = false shifter.enabled = false charliecloud.enabled = false + apptainer.enabled = false } docker { docker.enabled = true + docker.registry = 'quay.io' docker.userEmulation = true + conda.enabled = false singularity.enabled = false podman.enabled = false shifter.enabled = false charliecloud.enabled = false + apptainer.enabled = false } arm { docker.runOptions = '-u $(id -u):$(id -g) --platform=linux/amd64' @@ -98,31 +114,49 @@ profiles { singularity { singularity.enabled = true singularity.autoMounts = true + conda.enabled = false docker.enabled = false podman.enabled = false shifter.enabled = false charliecloud.enabled = false + apptainer.enabled = false } podman { podman.enabled = true + podman.registry = 'quay.io' + conda.enabled = false docker.enabled = false singularity.enabled = false shifter.enabled = false charliecloud.enabled = false + apptainer.enabled = false } shifter { shifter.enabled = true + conda.enabled = false docker.enabled = false singularity.enabled = false podman.enabled = false charliecloud.enabled = false + apptainer.enabled = false } charliecloud { charliecloud.enabled = true + conda.enabled = false + docker.enabled = false + singularity.enabled = false + podman.enabled = false + shifter.enabled = false + apptainer.enabled = false + } + apptainer { + apptainer.enabled = true + conda.enabled = false docker.enabled = false singularity.enabled = false podman.enabled = false shifter.enabled = false + charliecloud.enabled = false } gitpod { executor.name = 'local' @@ -169,12 +203,12 @@ dag { manifest { name = 'sanger-tol/variantcalling' - author = """Guoying Qi""" + author = """@muffato, @gq1, @priyanka-surana""" homePage = 'https://github.com/sanger-tol/variantcalling' - description = """variant calling""" + description = """Variant calling pipeline for PacBio data using DeepVariant""" mainScript = 'main.nf' nextflowVersion = '!>=22.10.1' - version = '1.0dev' + version = '1.0.0' doi = '' } diff --git a/nextflow_schema.json b/nextflow_schema.json index 73c70ad..cc5a4a1 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -1,8 +1,8 @@ { "$schema": "http://json-schema.org/draft-07/schema", - "$id": "https://raw.githubusercontent.com/sanger-tol/variantcalling/main/nextflow_schema.json", + "$id": "https://raw.githubusercontent.com/sanger-tol/variantcalling/master/nextflow_schema.json", "title": "sanger-tol/variantcalling pipeline parameters", - "description": "variant calling", + "description": "Variant calling pipeline for PacBio data using DeepVariant", "type": "object", "definitions": { "input_output_options": { @@ -34,6 +34,11 @@ "fa_icon": "fas fa-envelope", "help_text": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" + }, + "multiqc_title": { + "type": "string", + "description": "MultiQC report title. Printed as page header, used for filename if not otherwise specified.", + "fa_icon": "fas fa-file-signature" } } }, @@ -169,6 +174,14 @@ "fa_icon": "fas fa-remove-format", "hidden": true }, + "max_multiqc_email_size": { + "type": "string", + "description": "File size limit when attaching MultiQC reports to summary emails.", + "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$", + "default": "25.MB", + "fa_icon": "fas fa-file-upload", + "hidden": true + }, "monochrome_logs": { "type": "boolean", "description": "Do not use coloured log outputs.", @@ -182,6 +195,23 @@ "help_text": "Incoming hook URL for messaging service. Currently, MS Teams and Slack are supported.", "hidden": true }, + "multiqc_config": { + "type": "string", + "description": "Custom config file to supply to MultiQC.", + "fa_icon": "fas fa-cog", + "hidden": true + }, + "multiqc_logo": { + "type": "string", + "description": "Custom logo file to supply to MultiQC. File name must also be set in the MultiQC config file", + "fa_icon": "fas fa-image", + "hidden": true + }, + "multiqc_methods_description": { + "type": "string", + "description": "Custom MultiQC yaml file containing HTML including a methods description.", + "fa_icon": "fas fa-cog" + }, "tracedir": { "type": "string", "description": "Directory to keep pipeline Nextflow logs and reports.", diff --git a/pipeline_template.yml b/pipeline_template.yml index 0aa7398..09d0664 100644 --- a/pipeline_template.yml +++ b/pipeline_template.yml @@ -1,3 +1,3 @@ prefix: sanger-tol skip: - - igenomes +- igenomes diff --git a/tower.yml b/tower.yml new file mode 100644 index 0000000..787aedf --- /dev/null +++ b/tower.yml @@ -0,0 +1,5 @@ +reports: + multiqc_report.html: + display: "MultiQC HTML report" + samplesheet.csv: + display: "Auto-created samplesheet with collated metadata and FASTQ paths" diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index da91d34..59c0a7a 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -11,7 +11,7 @@ WorkflowVariantcalling.initialise(params, log) // TODO nf-core: Add all file path parameters for the pipeline to the list below // Check input path parameters to see if they exist -def checkPathParamList = [ params.input, params.fasta ] +def checkPathParamList = [ params.input, params.multiqc_config, params.fasta ] for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } // Check mandatory parameters @@ -23,6 +23,11 @@ if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input sample ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ +ch_multiqc_config = Channel.fromPath("$projectDir/assets/multiqc_config.yml", checkIfExists: true) +ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath( params.multiqc_config, checkIfExists: true ) : Channel.empty() +ch_multiqc_logo = params.multiqc_logo ? Channel.fromPath( params.multiqc_logo, checkIfExists: true ) : Channel.empty() +ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description, checkIfExists: true) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT LOCAL MODULES/SUBWORKFLOWS @@ -43,6 +48,8 @@ include { INPUT_CHECK } from '../subworkflows/local/input_check' // // MODULE: Installed directly from nf-core/modules // +include { FASTQC } from '../modules/nf-core/fastqc/main' +include { MULTIQC } from '../modules/nf-core/multiqc/main' include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' /* @@ -52,6 +59,7 @@ include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoft */ // Info required for completion email and summary +def multiqc_report = [] workflow VARIANTCALLING { @@ -65,10 +73,40 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) + // + // MODULE: Run FastQC + // + FASTQC ( + INPUT_CHECK.out.reads + ) + ch_versions = ch_versions.mix(FASTQC.out.versions.first()) + CUSTOM_DUMPSOFTWAREVERSIONS ( ch_versions.unique().collectFile(name: 'collated_versions.yml') ) + // + // MODULE: MultiQC + // + workflow_summary = WorkflowVariantcalling.paramsSummaryMultiqc(workflow, summary_params) + ch_workflow_summary = Channel.value(workflow_summary) + + methods_description = WorkflowVariantcalling.methodsDescriptionText(workflow, ch_multiqc_custom_methods_description) + ch_methods_description = Channel.value(methods_description) + + ch_multiqc_files = Channel.empty() + ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) + ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml')) + ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) + ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([])) + + MULTIQC ( + ch_multiqc_files.collect(), + ch_multiqc_config.toList(), + ch_multiqc_custom_config.toList(), + ch_multiqc_logo.toList() + ) + multiqc_report = MULTIQC.out.report.toList() } /* @@ -79,7 +117,7 @@ workflow VARIANTCALLING { workflow.onComplete { if (params.email || params.email_on_fail) { - NfcoreTemplate.email(workflow, params, summary_params, projectDir, log) + NfcoreTemplate.email(workflow, params, summary_params, projectDir, log, multiqc_report) } NfcoreTemplate.summary(workflow, params, log) if (params.hook_url) { From e54a47afa9b483894e65383b0f9208b65b393fe7 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 17:33:41 +0100 Subject: [PATCH 023/123] update the version to 1.1.0-dev --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 2363efa..8dba08f 100644 --- a/nextflow.config +++ b/nextflow.config @@ -209,7 +209,7 @@ manifest { description = """Variant calling pipeline for PacBio data using DeepVariant""" mainScript = 'main.nf' nextflowVersion = '!>=22.10.1' - version = '1.0.0' + version = '1.1.0-dev' doi = 'https://doi.org/10.5281/zenodo.7890527' } From 7d65f86aa940816f998b500e8fbc8abf1cf417eb Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 17:37:04 +0100 Subject: [PATCH 024/123] missing bracks --- lib/WorkflowVariantcalling.groovy | 1 + 1 file changed, 1 insertion(+) diff --git a/lib/WorkflowVariantcalling.groovy b/lib/WorkflowVariantcalling.groovy index 1b0fe03..d05a861 100755 --- a/lib/WorkflowVariantcalling.groovy +++ b/lib/WorkflowVariantcalling.groovy @@ -17,3 +17,4 @@ class WorkflowVariantcalling { Nextflow.error "Genome fasta file not specified with e.g. '--fasta genome.fa' or via a detectable config file." } } +} From 20062b4315a27dbb8a0dd763f9a342c547564817 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 17:41:53 +0100 Subject: [PATCH 025/123] update the workflow because deepvaritant module updated. --- subworkflows/local/deepvariant_caller.nf | 14 +++++++++++--- 1 file changed, 11 insertions(+), 3 deletions(-) diff --git a/subworkflows/local/deepvariant_caller.nf b/subworkflows/local/deepvariant_caller.nf index 08adb0f..10b5359 100644 --- a/subworkflows/local/deepvariant_caller.nf +++ b/subworkflows/local/deepvariant_caller.nf @@ -22,13 +22,21 @@ workflow DEEPVARIANT_CALLER { .set { cram_crai } // fasta - fasta = reads_fasta.map { meta, cram, crai, interval, fasta_file_name, fasta, fai -> [ fasta ] } + fasta = reads_fasta.map { meta, cram, crai, interval, fasta_file_name, fasta, fai -> + [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.type ], + fasta + ] + } // fai - fai = reads_fasta.map{ meta, cram, crai, interval, fasta_file_name, fasta, fai -> [ fai ] } + fai = reads_fasta.map{ meta, cram, crai, interval, fasta_file_name, fasta, fai -> + [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.type ], + fai + ] + } // split fasta in compressed format, no gzi index file needed - gzi = [] + gzi = [ [], [] ] // call deepvariant DEEPVARIANT ( cram_crai, fasta, fai, gzi ) From 9a8be9030d48d87f8509d5a2784bd7f678b67d98 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 17:44:46 +0100 Subject: [PATCH 026/123] nf-core modules update samtools/view --- modules.json | 2 +- modules/nf-core/samtools/view/main.nf | 4 ++-- modules/nf-core/samtools/view/meta.yml | 13 +++++++++---- 3 files changed, 12 insertions(+), 7 deletions(-) diff --git a/modules.json b/modules.json index 297798d..5d394c6 100644 --- a/modules.json +++ b/modules.json @@ -32,7 +32,7 @@ }, "samtools/view": { "branch": "master", - "git_sha": "371eff7748d769c2ddc8bd593773523a364a52fe", + "git_sha": "3ffae3598260a99e8db3207dead9f73f87f90d1f", "installed_by": ["modules"] } } diff --git a/modules/nf-core/samtools/view/main.nf b/modules/nf-core/samtools/view/main.nf index d7b2a0d..cb91fac 100644 --- a/modules/nf-core/samtools/view/main.nf +++ b/modules/nf-core/samtools/view/main.nf @@ -5,11 +5,11 @@ process SAMTOOLS_VIEW { conda "bioconda::samtools=1.17" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : - 'quay.io/biocontainers/samtools:1.17--h00cdaf9_0' }" + 'biocontainers/samtools:1.17--h00cdaf9_0' }" input: tuple val(meta), path(input), path(index) - path fasta + tuple val(meta2), path(fasta) path qname output: diff --git a/modules/nf-core/samtools/view/meta.yml b/modules/nf-core/samtools/view/meta.yml index 7691603..3b05450 100644 --- a/modules/nf-core/samtools/view/meta.yml +++ b/modules/nf-core/samtools/view/meta.yml @@ -26,12 +26,17 @@ input: description: BAM/CRAM/SAM file pattern: "*.{bam,cram,sam}" - index: - type: optional file - description: BAM.BAI/BAM.CSI/CRAM.CRAI file + type: file + description: BAM.BAI/BAM.CSI/CRAM.CRAI file (optional) pattern: "*.{.bai,.csi,.crai}" + - meta2: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'test' ] - fasta: - type: optional file - description: Reference file the CRAM was created with + type: file + description: Reference file the CRAM was created with (optional) pattern: "*.{fasta,fa}" - qname: type: file From a4f703370c9ec6b2553394c4b4b67feb65026676 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 17:57:54 +0100 Subject: [PATCH 027/123] update samtools view workflow because module updated --- subworkflows/local/input_filter_split.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/input_filter_split.nf b/subworkflows/local/input_filter_split.nf index f2f26c5..5963d36 100644 --- a/subworkflows/local/input_filter_split.nf +++ b/subworkflows/local/input_filter_split.nf @@ -62,7 +62,7 @@ workflow INPUT_FILTER_SPLIT { .set { fasta_fai } // filter reads - SAMTOOLS_VIEW ( reads, fasta, [] ) + SAMTOOLS_VIEW ( reads, [ [], fasta ], [] ) ch_versions = ch_versions.mix ( SAMTOOLS_VIEW.out.versions.first() ) // combine reads with splitted references From 048d08f867df6e4d05dfe5614fe1846f73ae9c7b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 17:59:18 +0100 Subject: [PATCH 028/123] nf-core modules update samtools/faidx --- modules.json | 2 +- modules/nf-core/samtools/faidx/main.nf | 18 ++++++++++++------ modules/nf-core/samtools/faidx/meta.yml | 14 ++++++++++++-- 3 files changed, 25 insertions(+), 9 deletions(-) diff --git a/modules.json b/modules.json index 5d394c6..bc363d0 100644 --- a/modules.json +++ b/modules.json @@ -27,7 +27,7 @@ }, "samtools/faidx": { "branch": "master", - "git_sha": "371eff7748d769c2ddc8bd593773523a364a52fe", + "git_sha": "fd742419940e01ba1c5ecb172c3e32ec840662fe", "installed_by": ["modules"] }, "samtools/view": { diff --git a/modules/nf-core/samtools/faidx/main.nf b/modules/nf-core/samtools/faidx/main.nf index 21be8ba..59ed308 100644 --- a/modules/nf-core/samtools/faidx/main.nf +++ b/modules/nf-core/samtools/faidx/main.nf @@ -5,15 +5,17 @@ process SAMTOOLS_FAIDX { conda "bioconda::samtools=1.17" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : - 'quay.io/biocontainers/samtools:1.17--h00cdaf9_0' }" + 'biocontainers/samtools:1.17--h00cdaf9_0' }" input: tuple val(meta), path(fasta) + tuple val(meta2), path(fai) output: - tuple val(meta), path ("*.fai"), emit: fai - tuple val(meta), path ("*.gzi"), emit: gzi, optional: true - path "versions.yml" , emit: versions + tuple val(meta), path ("*.{fa,fasta}") , emit: fa , optional: true + tuple val(meta), path ("*.fai") , emit: fai, optional: true + tuple val(meta), path ("*.gzi") , emit: gzi, optional: true + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when @@ -23,8 +25,8 @@ process SAMTOOLS_FAIDX { """ samtools \\ faidx \\ - $args \\ - $fasta + $fasta \\ + $args cat <<-END_VERSIONS > versions.yml "${task.process}": @@ -33,8 +35,12 @@ process SAMTOOLS_FAIDX { """ stub: + def match = (task.ext.args =~ /-o(?:utput)?\s(.*)\s?/).findAll() + def fastacmd = match[0] ? "touch ${match[0][1]}" : '' """ + ${fastacmd} touch ${fasta}.fai + cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/nf-core/samtools/faidx/meta.yml b/modules/nf-core/samtools/faidx/meta.yml index fe2fe9a..957b25e 100644 --- a/modules/nf-core/samtools/faidx/meta.yml +++ b/modules/nf-core/samtools/faidx/meta.yml @@ -3,6 +3,7 @@ description: Index FASTA file keywords: - index - fasta + - faidx tools: - samtools: description: | @@ -17,12 +18,21 @@ input: - meta: type: map description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] + Groovy Map containing reference information + e.g. [ id:'test' ] - fasta: type: file description: FASTA file pattern: "*.{fa,fasta}" + - meta2: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'test' ] + - fai: + type: file + description: FASTA index file + pattern: "*.{fai}" output: - meta: type: map From 555bc3da001b61bed56a576146db59ad0d6ddafe Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 18:19:41 +0100 Subject: [PATCH 029/123] update workflow because samtools faidx module updated --- subworkflows/local/input_filter_split.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/input_filter_split.nf b/subworkflows/local/input_filter_split.nf index 5963d36..c820901 100644 --- a/subworkflows/local/input_filter_split.nf +++ b/subworkflows/local/input_filter_split.nf @@ -49,7 +49,7 @@ workflow INPUT_FILTER_SPLIT { .set { split_fasta } // index split fasta files - SAMTOOLS_FAIDX ( split_fasta ) + SAMTOOLS_FAIDX ( split_fasta, [[], []]) ch_versions = ch_versions.mix( SAMTOOLS_FAIDX.out.versions.first() ) // join fasta with corresponding fai file From 6c4804be03d4b71348859b149ea1fedb10429f3b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 18:23:01 +0100 Subject: [PATCH 030/123] use version 2.8 for nf-core lint in github --- .github/workflows/linting.yml | 2 +- .nf-core.yml | 1 + 2 files changed, 2 insertions(+), 1 deletion(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 888cb4b..c20e72b 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -84,7 +84,7 @@ jobs: - name: Install dependencies run: | python -m pip install --upgrade pip - pip install nf-core + pip install nf-core==2.8.0 - name: Run nf-core lint env: diff --git a/.nf-core.yml b/.nf-core.yml index f58cf3f..4e011f7 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -16,6 +16,7 @@ lint: - lib/NfcoreTemplate.groovy - .github/PULL_REQUEST_TEMPLATE.md - .github/workflows/branch.yml + - .github/workflows/linting.yml - LICENSE - assets/email_template.html - .github/ISSUE_TEMPLATE/bug_report.yml From 07716c2cb48c76c29fad7e9af23ee1ab546d1961 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 18:30:46 +0100 Subject: [PATCH 031/123] remove white space --- lib/WorkflowVariantcalling.groovy | 1 - 1 file changed, 1 deletion(-) diff --git a/lib/WorkflowVariantcalling.groovy b/lib/WorkflowVariantcalling.groovy index d05a861..523a779 100755 --- a/lib/WorkflowVariantcalling.groovy +++ b/lib/WorkflowVariantcalling.groovy @@ -11,7 +11,6 @@ class WorkflowVariantcalling { // Check and validate parameters // public static void initialise(params, log) { - if (!params.fasta) { Nextflow.error "Genome fasta file not specified with e.g. '--fasta genome.fa' or via a detectable config file." From 9e731d7061d7aaa892436f6e4472127e49429381 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 18:37:19 +0100 Subject: [PATCH 032/123] prettier --- .github/workflows/linting.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index c20e72b..3f27dab 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -84,7 +84,7 @@ jobs: - name: Install dependencies run: | python -m pip install --upgrade pip - pip install nf-core==2.8.0 + pip install nf-core==2.8.0 - name: Run nf-core lint env: From b302172850a721b98238f84d842c987389b24c2c Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 21 Sep 2023 18:53:40 +0100 Subject: [PATCH 033/123] to fail prettier --- .github/workflows/linting.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 3f27dab..c20e72b 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -84,7 +84,7 @@ jobs: - name: Install dependencies run: | python -m pip install --upgrade pip - pip install nf-core==2.8.0 + pip install nf-core==2.8.0 - name: Run nf-core lint env: From d50080ef2750b01fe5da3af9a031e286fcf571b4 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Thu, 21 Sep 2023 17:54:47 +0000 Subject: [PATCH 034/123] [automated] Fix linting with Prettier --- .github/workflows/linting.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index c20e72b..3f27dab 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -84,7 +84,7 @@ jobs: - name: Install dependencies run: | python -m pip install --upgrade pip - pip install nf-core==2.8.0 + pip install nf-core==2.8.0 - name: Run nf-core lint env: From f5a1186549dc155bad81622202462bdb15ebe2d1 Mon Sep 17 00:00:00 2001 From: Priyanka Surana Date: Tue, 26 Sep 2023 17:24:42 +0100 Subject: [PATCH 035/123] Update nextflow.config --- nextflow.config | 1 + 1 file changed, 1 insertion(+) diff --git a/nextflow.config b/nextflow.config index 8dba08f..83e274f 100644 --- a/nextflow.config +++ b/nextflow.config @@ -115,6 +115,7 @@ profiles { singularity { singularity.enabled = true singularity.autoMounts = true + singularity.registry = 'quay.io' conda.enabled = false docker.enabled = false podman.enabled = false From fe45c30046d4d0eaaf30a0ea156c881035919e9d Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 26 Sep 2023 17:27:11 +0100 Subject: [PATCH 036/123] update the display text. --- tower.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/tower.yml b/tower.yml index c61323c..48611ab 100644 --- a/tower.yml +++ b/tower.yml @@ -1,3 +1,3 @@ reports: samplesheet.csv: - display: "Auto-created samplesheet with collated metadata and FASTQ paths" + display: "Auto-created samplesheet with collated metadata and aligned file paths." From 26e7b3dea0463efae8c650403bf1d8876ab8eccc Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 29 Sep 2023 14:53:46 +0100 Subject: [PATCH 037/123] remove unused log params --- lib/WorkflowMain.groovy | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index f568e77..0309eb4 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -22,7 +22,7 @@ class WorkflowMain { // // Generate help string // - public static String help(workflow, params, log) { + public static String help(workflow, params) { def command = "nextflow run ${workflow.manifest.name} --input samplesheet.csv --fasta reference.fa --fai reference.fai --outdir results -profile docker" def help_string = '' help_string += NfcoreTemplate.logo(workflow, params.monochrome_logs) From 3f89203dcc7c1182b9f784c17e65cc1b6d9fcbfd Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 14:15:19 +0100 Subject: [PATCH 038/123] Remove index file from the samplesheet and update checking script --- assets/samplesheet.csv | 9 +++++---- assets/samplesheet_test.csv | 9 +++++---- assets/samplesheet_test_full.csv | 4 ++-- assets/schema_input.json | 7 +------ bin/check_samplesheet.py | 29 +++++++---------------------- 5 files changed, 20 insertions(+), 38 deletions(-) diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv index 2ea95db..9de2e5b 100644 --- a/assets/samplesheet.csv +++ b/assets/samplesheet.csv @@ -1,4 +1,5 @@ -sample,datatype,datafile,indexfile -sample1,pacbio,/path/to/data/file/file1.bam,/path/to/index/file/file1.bam.bai -sample2,pacbio,/path/to/data/file/file2.cram,/path/to/index/file/file2.cram.crai -sample3,pacbio,/path/to/data/file/file3.bam,/path/to/index/file/file3.bam.csi +sample,datatype,datafile +sample1,pacbio,/path/to/data/file/file1.bam +sample2,pacbio,/path/to/data/file/file2.cram +sample3,pacbio,/path/to/data/file/file3-1.bam +sample3,pacbio,/path/to/data/file/file3-2.cram diff --git a/assets/samplesheet_test.csv b/assets/samplesheet_test.csv index cf5546a..6eb03e5 100644 --- a/assets/samplesheet_test.csv +++ b/assets/samplesheet_test.csv @@ -1,4 +1,5 @@ -sample,datatype,datafile,indexfile -icCanRufa1_crai,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.cram,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.cram.crai -icCanRufa1_bai,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bam,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bam.bai -icCanRufa1_csi,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bam,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bam.csi +sample,datatype,datafile +icCanRufa1_cram,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.cram +icCanRufa1_bam,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bam +icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.cram +icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bam diff --git a/assets/samplesheet_test_full.csv b/assets/samplesheet_test_full.csv index 4495dff..1e40e2b 100644 --- a/assets/samplesheet_test_full.csv +++ b/assets/samplesheet_test_full.csv @@ -1,2 +1,2 @@ -sample,datatype,datafile,indexfile -icCanRufa1,pacbio,/lustre/scratch123/tol/resources/nextflow/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram,/lustre/scratch123/tol/resources/nextflow/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram.crai +sample,datatype,datafile +icCanRufa1,pacbio,/lustre/scratch123/tol/resources/nextflow/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1.cram diff --git a/assets/schema_input.json b/assets/schema_input.json index 43497e9..f264cf6 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -21,13 +21,8 @@ "type": "string", "pattern": "^\\S+\\.(bam|cram)$", "errorMessage": "Data file for reads cannot contain spaces and must have extension 'cram' or 'bam'" - }, - "indexfile": { - "type": "string", - "pattern": "^\\S+\\.(bai|csi|crai)$", - "errorMessage": "Data index file for reads cannot contain spaces and must have extension 'bai', 'csi' or 'crai'" } }, - "required": ["sample", "datatype", "datafile", "indexfile"] + "required": ["sample", "datatype", "datafile"] } } diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py index 6bbd806..52af146 100755 --- a/bin/check_samplesheet.py +++ b/bin/check_samplesheet.py @@ -35,7 +35,6 @@ def __init__( sample_col="sample", type_col="datatype", file_col="datafile", - index_col="indexfile", **kwargs, ): """ @@ -48,8 +47,6 @@ def __init__( the read data (default "datatype"). file_col (str): The name of the column that contains the file path for the read data (default "datafile"). - index_col (str): The name of the column that contains the index file - for the data (default "indexfile"). """ super().__init__(**kwargs) @@ -57,7 +54,6 @@ def __init__( self._sample_col = sample_col self._type_col = type_col self._file_col = file_col - self._index_col = index_col self._seen = set() self.modified = [] @@ -73,7 +69,6 @@ def validate_and_transform(self, row): self._validate_sample(row) self._validate_type(row) self._validate_data_file(row) - self._validate_index_file(row) self._seen.add((row[self._sample_col], row[self._file_col])) self.modified.append(row) @@ -98,17 +93,6 @@ def _validate_data_file(self, row): raise AssertionError("Data file is required.") self._validate_data_format(row[self._file_col]) - def _validate_index_file(self, row): - """Assert that the indexfile is non-empty and has the right format.""" - if len(row[self._index_col]) <= 0: - raise AssertionError("Data index file is required.") - if row[self._file_col].endswith("bam") and not ( - row[self._index_col].endswith("bai") or row[self._index_col].endswith("csi") - ): - raise AssertionError("bai or csi index file should be given for bam file.") - if row[self._file_col].endswith("cram") and not row[self._index_col].endswith("crai"): - raise AssertionError("crai index file shuld be given for cram file.") - def _validate_data_format(self, filename): """Assert that a given filename has one of the expected read data file extensions.""" if not any(filename.endswith(extension) for extension in self.DATA_VALID_FORMATS): @@ -162,7 +146,7 @@ def sniff_format(handle): peek = read_head(handle) handle.seek(0) sniffer = csv.Sniffer() - # same input file could retrun random true or false + # same input file could return random true or false # disable it now # the following validation should be enough # if not sniffer.has_header(peek): @@ -188,16 +172,17 @@ def check_samplesheet(file_in, file_out): This function checks that the samplesheet follows the following structure, see also the `variantcalling samplesheet`_:: - sample,datatype,datafile,indexfile - sample1,pacbio,/path/to/data/file/file1.bam,/path/to/index/file/file1.bam.bai - sample2,pacbio,/path/to/data/file/file2.cram,/path/to/index/file/file2.cram.crai - sample3,pacbio,/path/to/data/file/file3.bam,/path/to/index/file/file3.bam.csi + sample,datatype,datafile + sample1,pacbio,/path/to/data/file/file1.bam + sample2,pacbio,/path/to/data/file/file2.cram + sample3,pacbio,/path/to/data/file/file3-1.bam + sample3,pacbio,/path/to/data/file/file3-2.cram .. _variantcalling samplesheet: https://raw.githubusercontent.com/sanger-tol/variantcalling/main/assets/samplesheet.csv """ - required_columns = {"sample", "datatype", "datafile", "indexfile"} + required_columns = {"sample", "datatype", "datafile"} # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. with file_in.open(newline="") as in_handle: reader = csv.DictReader(in_handle, dialect=sniff_format(in_handle)) From cef527b538f5c6e72d014cd7f9eae9c0eec584dc Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 14:50:54 +0100 Subject: [PATCH 039/123] Remove indexfile from Input_check workflow --- subworkflows/local/input_check.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index d2f72e9..30a07d5 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -12,10 +12,10 @@ workflow INPUT_CHECK { SAMPLESHEET_CHECK ( samplesheet ) .csv .splitCsv ( header:true, sep:',' ) - .map { [[id: it.sample, type: it.datatype], file(it.datafile), file(it.indexfile)] } + .map { [[id: it.sample, type: it.datatype], file(it.datafile)] } .set { reads } emit: - reads // channel: [ val(meta), data, index ] + reads // channel: [ val(meta), data ] versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ] } From 67826e3eeaaa6a7007b562103b72551f83332f9e Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 17:42:56 +0100 Subject: [PATCH 040/123] add sample name to the meta data --- subworkflows/local/input_check.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index 30a07d5..aa0bbc9 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -12,9 +12,9 @@ workflow INPUT_CHECK { SAMPLESHEET_CHECK ( samplesheet ) .csv .splitCsv ( header:true, sep:',' ) - .map { [[id: it.sample, type: it.datatype], file(it.datafile)] } + .map { [[id: it.sample, sample: it.sample.replaceAll(/_T\d+$/, ''), type: it.datatype], file(it.datafile)] } .set { reads } - + emit: reads // channel: [ val(meta), data ] versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ] From 2d30d826a2a48bece2af926d00d93a672aa1e47d Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 17:55:56 +0100 Subject: [PATCH 041/123] nf-core modules install samtools/merge --- modules.json | 5 ++ modules/nf-core/samtools/merge/main.nf | 56 +++++++++++++++++++ modules/nf-core/samtools/merge/meta.yml | 73 +++++++++++++++++++++++++ 3 files changed, 134 insertions(+) create mode 100644 modules/nf-core/samtools/merge/main.nf create mode 100644 modules/nf-core/samtools/merge/meta.yml diff --git a/modules.json b/modules.json index bc363d0..6f79373 100644 --- a/modules.json +++ b/modules.json @@ -30,6 +30,11 @@ "git_sha": "fd742419940e01ba1c5ecb172c3e32ec840662fe", "installed_by": ["modules"] }, + "samtools/merge": { + "branch": "master", + "git_sha": "0460d316170f75f323111b4a2c0a2989f0c32013", + "installed_by": ["modules"] + }, "samtools/view": { "branch": "master", "git_sha": "3ffae3598260a99e8db3207dead9f73f87f90d1f", diff --git a/modules/nf-core/samtools/merge/main.nf b/modules/nf-core/samtools/merge/main.nf new file mode 100644 index 0000000..b73b7cb --- /dev/null +++ b/modules/nf-core/samtools/merge/main.nf @@ -0,0 +1,56 @@ +process SAMTOOLS_MERGE { + tag "$meta.id" + label 'process_low' + + conda "bioconda::samtools=1.17" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(input_files, stageAs: "?/*") + tuple val(meta2), path(fasta) + tuple val(meta3), path(fai) + + output: + tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam + tuple val(meta), path("${prefix}.cram"), optional:true, emit: cram + tuple val(meta), path("*.csi") , optional:true, emit: csi + path "versions.yml" , emit: versions + + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + prefix = task.ext.prefix ?: "${meta.id}" + def file_type = input_files instanceof List ? input_files[0].getExtension() : input_files.getExtension() + def reference = fasta ? "--reference ${fasta}" : "" + """ + samtools \\ + merge \\ + --threads ${task.cpus-1} \\ + $args \\ + ${reference} \\ + ${prefix}.${file_type} \\ + $input_files + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ + + stub: + prefix = task.ext.suffix ? "${meta.id}${task.ext.suffix}" : "${meta.id}" + def file_type = input_files instanceof List ? input_files[0].getExtension() : input_files.getExtension() + """ + touch ${prefix}.${file_type} + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/merge/meta.yml b/modules/nf-core/samtools/merge/meta.yml new file mode 100644 index 0000000..3a815f7 --- /dev/null +++ b/modules/nf-core/samtools/merge/meta.yml @@ -0,0 +1,73 @@ +name: samtools_merge +description: Merge BAM or CRAM file +keywords: + - merge + - bam + - sam + - cram +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - input_files: + type: file + description: BAM/CRAM file + pattern: "*.{bam,cram,sam}" + - meta2: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'genome' ] + - fasta: + type: file + description: Reference file the CRAM was created with (optional) + pattern: "*.{fasta,fa}" + - meta3: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'genome' ] + - fai: + type: file + description: Index of the reference file the CRAM was created with (optional) + pattern: "*.fai" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: BAM file + pattern: "*.{bam}" + - cram: + type: file + description: CRAM file + pattern: "*.{cram}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - csi: + type: file + description: BAM index file (optional) + pattern: "*.csi" +authors: + - "@drpatelh" + - "@yuukiiwa " + - "@maxulysse" + - "@FriederikeHanssen" + - "@ramprasadn" From 953658f86209de21f23627f376f9d688ecc73acb Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 21:12:54 +0100 Subject: [PATCH 042/123] Add input merge sub workflow --- subworkflows/local/input_merge.nf | 44 +++++++++++++++++++++++++++++++ workflows/variantcalling.nf | 10 +++++++ 2 files changed, 54 insertions(+) create mode 100644 subworkflows/local/input_merge.nf diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf new file mode 100644 index 0000000..121c246 --- /dev/null +++ b/subworkflows/local/input_merge.nf @@ -0,0 +1,44 @@ +// +// Merge READS(bam or cram files) together by sample name +// + +include { SAMTOOLS_MERGE } from '../../modules/nf-core/samtools/merge' + +workflow INPUT_MERGE { + take: + fasta // file: /path/to/genome.fasta or /path/to/genome.fasta.gz + fai // file: /path/to/genome.*.fai + gzi // file: /path/to/genome.fasta.gz.gzi or null + reads // channel: [ val(meta), data ] + + main: + // group input reads file by sample name + reads + .map{ it -> [ it[0].sample, it[1] ] } + .groupTuple() + .set{ merged_reads } + + // group input meta data together by sample name as well + // use the first meta data for the combined reads + reads + .map{ it -> [ it[0].sample, it[0] ] } + .groupTuple() + .map { it -> [it[0], it[1][0]] } + .join( merged_reads ) + .map { it -> [ it[1] , it [2] ]} + .set { merged_reads_with_meta } + + // call samtool merge + SAMTOOLS_MERGE( merged_reads_with_meta, + [ [], fasta ], + [ [], fai ], + [ [], gzi ] + ) + + emit: + bam = SAMTOOLS_MERGE.out.bam + cram = SAMTOOLS_MERGE.out.cram + csi = SAMTOOLS_MERGE.out.csi + versions = SAMTOOLS_MERGE.out.versions // channel: [ versions.yml ] + +} diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 6c6ce09..de68f0b 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -47,6 +47,7 @@ if (params.split_fasta_cutoff ) { split_fasta_cutoff = params.split_fasta_cutoff // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' +include { INPUT_MERGE } from '../subworkflows/local/input_merge' include { INPUT_FILTER_SPLIT } from '../subworkflows/local/input_filter_split' include { DEEPVARIANT_CALLER } from '../subworkflows/local/deepvariant_caller' @@ -81,6 +82,15 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) + INPUT_MERGE ( + fasta_file, + fai_file, + gzi_file, + INPUT_CHECK.out.reads + ) + ch_versions = ch_versions.mix(INPUT_MERGE.out.versions) + + // // SUBWORKFLOW: split the input fasta file and filter input reads // From 0d0d06e80093fb9a81cab0ea60032c9d1434bb70 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 21:41:47 +0100 Subject: [PATCH 043/123] comments --- workflows/variantcalling.nf | 3 +++ 1 file changed, 3 insertions(+) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index de68f0b..5b2c581 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -82,6 +82,9 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) + // + // SUBWORKFLOW: merge the input reads by sample name + // INPUT_MERGE ( fasta_file, fai_file, From 224c7f23a0954875fdd1e10179e4d2c33dfa5b4f Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 5 Oct 2023 21:45:11 +0100 Subject: [PATCH 044/123] patch samtools_merge module to allow using fasta.gz file with gzi index file. --- modules.json | 3 ++- modules/nf-core/samtools/merge/main.nf | 1 + modules/nf-core/samtools/merge/meta.yml | 4 +++ .../samtools/merge/samtools-merge.diff | 27 +++++++++++++++++++ 4 files changed, 34 insertions(+), 1 deletion(-) create mode 100644 modules/nf-core/samtools/merge/samtools-merge.diff diff --git a/modules.json b/modules.json index 6f79373..133358e 100644 --- a/modules.json +++ b/modules.json @@ -33,7 +33,8 @@ "samtools/merge": { "branch": "master", "git_sha": "0460d316170f75f323111b4a2c0a2989f0c32013", - "installed_by": ["modules"] + "installed_by": ["modules"], + "patch": "modules/nf-core/samtools/merge/samtools-merge.diff" }, "samtools/view": { "branch": "master", diff --git a/modules/nf-core/samtools/merge/main.nf b/modules/nf-core/samtools/merge/main.nf index b73b7cb..3c7faf4 100644 --- a/modules/nf-core/samtools/merge/main.nf +++ b/modules/nf-core/samtools/merge/main.nf @@ -11,6 +11,7 @@ process SAMTOOLS_MERGE { tuple val(meta), path(input_files, stageAs: "?/*") tuple val(meta2), path(fasta) tuple val(meta3), path(fai) + tuple val(meta4), path(gzi) output: tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam diff --git a/modules/nf-core/samtools/merge/meta.yml b/modules/nf-core/samtools/merge/meta.yml index 3a815f7..4a6bc23 100644 --- a/modules/nf-core/samtools/merge/meta.yml +++ b/modules/nf-core/samtools/merge/meta.yml @@ -43,6 +43,10 @@ input: type: file description: Index of the reference file the CRAM was created with (optional) pattern: "*.fai" + - gzi: + type: file + description: Index of the reference file the CRAM was created with (optional) + pattern: "*.gzi" output: - meta: type: map diff --git a/modules/nf-core/samtools/merge/samtools-merge.diff b/modules/nf-core/samtools/merge/samtools-merge.diff new file mode 100644 index 0000000..4c48cc0 --- /dev/null +++ b/modules/nf-core/samtools/merge/samtools-merge.diff @@ -0,0 +1,27 @@ +Changes in module 'nf-core/samtools/merge' +--- modules/nf-core/samtools/merge/meta.yml ++++ modules/nf-core/samtools/merge/meta.yml +@@ -43,6 +43,10 @@ + type: file + description: Index of the reference file the CRAM was created with (optional) + pattern: "*.fai" ++ - gzi: ++ type: file ++ description: Index of the reference file the CRAM was created with (optional) ++ pattern: "*.gzi" + output: + - meta: + type: map + +--- modules/nf-core/samtools/merge/main.nf ++++ modules/nf-core/samtools/merge/main.nf +@@ -11,6 +11,7 @@ + tuple val(meta), path(input_files, stageAs: "?/*") + tuple val(meta2), path(fasta) + tuple val(meta3), path(fai) ++ tuple val(meta4), path(gzi) + + output: + tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam + +************************************************************ From 36f4c677e30bc4b630144d678bfc945fe3474f16 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 09:26:55 +0100 Subject: [PATCH 045/123] use original sample name for id if just 1, otherwise add _combined. --- subworkflows/local/input_merge.nf | 9 +++++++-- 1 file changed, 7 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 121c246..d2eafb5 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -25,7 +25,12 @@ workflow INPUT_MERGE { .groupTuple() .map { it -> [it[0], it[1][0]] } .join( merged_reads ) - .map { it -> [ it[1] , it [2] ]} + .map { it -> [ + [ id: ( it[2].size() == 1 ) ? it[1].sample : it[1].sample + '_combined', + type: it[1].type + ], + it[2] + ]} .set { merged_reads_with_meta } // call samtool merge @@ -38,7 +43,7 @@ workflow INPUT_MERGE { emit: bam = SAMTOOLS_MERGE.out.bam cram = SAMTOOLS_MERGE.out.cram - csi = SAMTOOLS_MERGE.out.csi + csi = SAMTOOLS_MERGE.out.csi versions = SAMTOOLS_MERGE.out.versions // channel: [ versions.yml ] } From a808cf24897fc3e8c3a87b35a55dcd99cd07faa4 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 10:16:31 +0100 Subject: [PATCH 046/123] Path samtools_merge module again add indexing, emit crai index file as well --- modules/nf-core/samtools/merge/main.nf | 2 ++ modules/nf-core/samtools/merge/meta.yml | 4 +++ .../samtools/merge/samtools-merge.diff | 27 ++++++++++++++++++- 3 files changed, 32 insertions(+), 1 deletion(-) diff --git a/modules/nf-core/samtools/merge/main.nf b/modules/nf-core/samtools/merge/main.nf index 3c7faf4..90ddfbe 100644 --- a/modules/nf-core/samtools/merge/main.nf +++ b/modules/nf-core/samtools/merge/main.nf @@ -17,6 +17,7 @@ process SAMTOOLS_MERGE { tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam tuple val(meta), path("${prefix}.cram"), optional:true, emit: cram tuple val(meta), path("*.csi") , optional:true, emit: csi + tuple val(meta), path("*.crai") , optional:true, emit: crai path "versions.yml" , emit: versions @@ -32,6 +33,7 @@ process SAMTOOLS_MERGE { samtools \\ merge \\ --threads ${task.cpus-1} \\ + --write-index \\ $args \\ ${reference} \\ ${prefix}.${file_type} \\ diff --git a/modules/nf-core/samtools/merge/meta.yml b/modules/nf-core/samtools/merge/meta.yml index 4a6bc23..056a95f 100644 --- a/modules/nf-core/samtools/merge/meta.yml +++ b/modules/nf-core/samtools/merge/meta.yml @@ -69,6 +69,10 @@ output: type: file description: BAM index file (optional) pattern: "*.csi" + - crai: + type: file + description: CRAM index file (optional) + pattern: "*.crai" authors: - "@drpatelh" - "@yuukiiwa " diff --git a/modules/nf-core/samtools/merge/samtools-merge.diff b/modules/nf-core/samtools/merge/samtools-merge.diff index 4c48cc0..a740ce0 100644 --- a/modules/nf-core/samtools/merge/samtools-merge.diff +++ b/modules/nf-core/samtools/merge/samtools-merge.diff @@ -12,10 +12,21 @@ Changes in module 'nf-core/samtools/merge' output: - meta: type: map +@@ -65,6 +69,10 @@ + type: file + description: BAM index file (optional) + pattern: "*.csi" ++ - crai: ++ type: file ++ description: CRAM index file (optional) ++ pattern: "*.crai" + authors: + - "@drpatelh" + - "@yuukiiwa " --- modules/nf-core/samtools/merge/main.nf +++ modules/nf-core/samtools/merge/main.nf -@@ -11,6 +11,7 @@ +@@ -11,11 +11,13 @@ tuple val(meta), path(input_files, stageAs: "?/*") tuple val(meta2), path(fasta) tuple val(meta3), path(fai) @@ -23,5 +34,19 @@ Changes in module 'nf-core/samtools/merge' output: tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam + tuple val(meta), path("${prefix}.cram"), optional:true, emit: cram + tuple val(meta), path("*.csi") , optional:true, emit: csi ++ tuple val(meta), path("*.crai") , optional:true, emit: crai + path "versions.yml" , emit: versions + + +@@ -31,6 +33,7 @@ + samtools \\ + merge \\ + --threads ${task.cpus-1} \\ ++ --write-index \\ + $args \\ + ${reference} \\ + ${prefix}.${file_type} \\ ************************************************************ From c646b8b1b4b4e5b979347f9f625f89c7e6ca6e01 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 10:27:29 +0100 Subject: [PATCH 047/123] emit crai files as well --- subworkflows/local/input_merge.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index d2eafb5..a0e7220 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -44,6 +44,7 @@ workflow INPUT_MERGE { bam = SAMTOOLS_MERGE.out.bam cram = SAMTOOLS_MERGE.out.cram csi = SAMTOOLS_MERGE.out.csi + crai = SAMTOOLS_MERGE.out.crai versions = SAMTOOLS_MERGE.out.versions // channel: [ versions.yml ] } From c4a762fed89d13c9e39ee99c017d30e4a1239ce0 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 10:43:26 +0100 Subject: [PATCH 048/123] nf-core modules install samtools/sort --- modules.json | 5 +++ modules/nf-core/samtools/sort/main.nf | 49 ++++++++++++++++++++++++++ modules/nf-core/samtools/sort/meta.yml | 48 +++++++++++++++++++++++++ 3 files changed, 102 insertions(+) create mode 100644 modules/nf-core/samtools/sort/main.nf create mode 100644 modules/nf-core/samtools/sort/meta.yml diff --git a/modules.json b/modules.json index 133358e..183f5b3 100644 --- a/modules.json +++ b/modules.json @@ -36,6 +36,11 @@ "installed_by": ["modules"], "patch": "modules/nf-core/samtools/merge/samtools-merge.diff" }, + "samtools/sort": { + "branch": "master", + "git_sha": "a0f7be95788366c1923171e358da7d049eb440f9", + "installed_by": ["modules"] + }, "samtools/view": { "branch": "master", "git_sha": "3ffae3598260a99e8db3207dead9f73f87f90d1f", diff --git a/modules/nf-core/samtools/sort/main.nf b/modules/nf-core/samtools/sort/main.nf new file mode 100644 index 0000000..2b7753f --- /dev/null +++ b/modules/nf-core/samtools/sort/main.nf @@ -0,0 +1,49 @@ +process SAMTOOLS_SORT { + tag "$meta.id" + label 'process_medium' + + conda "bioconda::samtools=1.17" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(bam) + + output: + tuple val(meta), path("*.bam"), emit: bam + tuple val(meta), path("*.csi"), emit: csi, optional: true + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + if ("$bam" == "${prefix}.bam") error "Input and output names are the same, use \"task.ext.prefix\" to disambiguate!" + """ + samtools sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/sort/meta.yml b/modules/nf-core/samtools/sort/meta.yml new file mode 100644 index 0000000..0732843 --- /dev/null +++ b/modules/nf-core/samtools/sort/meta.yml @@ -0,0 +1,48 @@ +name: samtools_sort +description: Sort SAM/BAM/CRAM file +keywords: + - sort + - bam + - sam + - cram +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: Sorted BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - csi: + type: file + description: BAM index file (optional) + pattern: "*.csi" +authors: + - "@drpatelh" + - "@ewels" From 9fa3696989b7d4398654c507e0018d983fb37067 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 11:37:33 +0100 Subject: [PATCH 049/123] Add an option to sort input if not sorted. --- nextflow.config | 1 + nextflow_schema.json | 4 ++++ subworkflows/local/input_merge.nf | 18 ++++++++++++++++-- workflows/variantcalling.nf | 6 ++++-- 4 files changed, 25 insertions(+), 4 deletions(-) diff --git a/nextflow.config b/nextflow.config index 83e274f..4659669 100644 --- a/nextflow.config +++ b/nextflow.config @@ -11,6 +11,7 @@ params { // Input options input = null + sort_input = false fasta = null fai = null gzi = null diff --git a/nextflow_schema.json b/nextflow_schema.json index 3a5272c..5e04d99 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -28,6 +28,10 @@ "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.", "fa_icon": "fas fa-folder-open" }, + "sort_input": { + "type": "boolean", + "description": "Boolean whether to sort input reads files" + }, "email": { "type": "string", "description": "Email address for completion summary.", diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index a0e7220..53b5e0f 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -3,6 +3,7 @@ // include { SAMTOOLS_MERGE } from '../../modules/nf-core/samtools/merge' +include { SAMTOOLS_SORT } from '../../modules/nf-core/samtools/sort' workflow INPUT_MERGE { take: @@ -10,10 +11,22 @@ workflow INPUT_MERGE { fai // file: /path/to/genome.*.fai gzi // file: /path/to/genome.fasta.gz.gzi or null reads // channel: [ val(meta), data ] + sort_input // bollean: true or false main: + ch_versions = Channel.empty() + + // sort input reads if asked + if ( sort_input ) { + SAMTOOLS_SORT( reads ) + ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions ) + sorted_reads = SAMTOOLS_SORT.out.bam + } else { + sorted_reads = reads + } + // group input reads file by sample name - reads + sorted_reads .map{ it -> [ it[0].sample, it[1] ] } .groupTuple() .set{ merged_reads } @@ -39,12 +52,13 @@ workflow INPUT_MERGE { [ [], fai ], [ [], gzi ] ) + ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions ) emit: bam = SAMTOOLS_MERGE.out.bam cram = SAMTOOLS_MERGE.out.cram csi = SAMTOOLS_MERGE.out.csi crai = SAMTOOLS_MERGE.out.crai - versions = SAMTOOLS_MERGE.out.versions // channel: [ versions.yml ] + versions = ch_versions // channel: [ versions.yml ] } diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 5b2c581..2c1a78f 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -28,7 +28,8 @@ if (params.gzi) { } // Check optional parameters -if (params.interval) { interval_file = file(params.interval) } else { interval_file = null } +if (params.sort_input) { sort_input = params.sort_input } else { sort_input = false } +if (params.interval) { interval_file = file(params.interval) } else { interval_file = null } if (params.split_fasta_cutoff ) { split_fasta_cutoff = params.split_fasta_cutoff } else { split_fasta_cutoff = 100000 } /* @@ -89,7 +90,8 @@ workflow VARIANTCALLING { fasta_file, fai_file, gzi_file, - INPUT_CHECK.out.reads + INPUT_CHECK.out.reads, + sort_input ) ch_versions = ch_versions.mix(INPUT_MERGE.out.versions) From 5cbea5bd50e0816ab7946c29d006d0975b6532c9 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 12:03:27 +0100 Subject: [PATCH 050/123] combine merged bam/cram together, add with their index files as well. --- subworkflows/local/input_merge.nf | 13 +++++++++---- 1 file changed, 9 insertions(+), 4 deletions(-) diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 53b5e0f..da9047e 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -54,11 +54,16 @@ workflow INPUT_MERGE { ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions ) + SAMTOOLS_MERGE.out.bam + .join(SAMTOOLS_MERGE.out.csi) + .concat( + SAMTOOLS_MERGE.out.cram + .join(SAMTOOLS_MERGE.out.crai) + ) + .set{ indexed_merged_reads }; + emit: - bam = SAMTOOLS_MERGE.out.bam - cram = SAMTOOLS_MERGE.out.cram - csi = SAMTOOLS_MERGE.out.csi - crai = SAMTOOLS_MERGE.out.crai + indexed_merged_reads = indexed_merged_reads versions = ch_versions // channel: [ versions.yml ] } From 3ac73e9482d849661920ccc216f0d2cb5ff3dcca Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 12:20:16 +0100 Subject: [PATCH 051/123] add filtered to distinguish the samtools input and output name --- conf/modules.config | 1 + 1 file changed, 1 insertion(+) diff --git a/conf/modules.config b/conf/modules.config index 7dc8677..6672bfc 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -22,6 +22,7 @@ process { withName: '.*:INPUT_FILTER_SPLIT:SAMTOOLS_VIEW' { ext.args = '--output-fmt cram --write-index -F 0x900' + ext.prefix = { "${meta.id}_filtered" } } withName: '.*:DEEPVARIANT_CALLER:DEEPVARIANT' { From bc6e5a0982503673e2898347b8d2253b21da6ea5 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 12:22:42 +0100 Subject: [PATCH 052/123] use the merged read for the rest of pipeline --- workflows/variantcalling.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 2c1a78f..3b36a99 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -103,7 +103,7 @@ workflow VARIANTCALLING { fasta_file, fai_file, gzi_file, - INPUT_CHECK.out.reads, + INPUT_MERGE.out.indexed_merged_reads, interval_file, split_fasta_cutoff ) From a7131a8d087c2e69e46ef9c0164d70e19310e916 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 19:00:58 +0100 Subject: [PATCH 053/123] covert all input files into channels, and make reference fasta index file optional --- conf/test.config | 4 +- nextflow_schema.json | 2 +- subworkflows/local/input_filter_split.nf | 10 ++-- subworkflows/local/input_merge.nf | 17 +++++-- workflows/variantcalling.nf | 60 +++++++++++++++--------- 5 files changed, 61 insertions(+), 32 deletions(-) diff --git a/conf/test.config b/conf/test.config index be32ebe..f89e185 100644 --- a/conf/test.config +++ b/conf/test.config @@ -26,8 +26,8 @@ params { fasta = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz' // Reference index file - fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.fai' - gzi = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' + // fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.fai' + // gzi = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' // Interval bed file interval = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bed' diff --git a/nextflow_schema.json b/nextflow_schema.json index 5e04d99..dc6428b 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -70,7 +70,7 @@ "description": "The minimum fasta file size when splitting the input fasta file by sequence." } }, - "required": ["fasta", "fai"] + "required": ["fasta"] }, "institutional_config_options": { "title": "Institutional config options", diff --git a/subworkflows/local/input_filter_split.nf b/subworkflows/local/input_filter_split.nf index c820901..95555d7 100644 --- a/subworkflows/local/input_filter_split.nf +++ b/subworkflows/local/input_filter_split.nf @@ -19,8 +19,7 @@ workflow INPUT_FILTER_SPLIT { ch_versions = Channel.empty() // split the fasta file into files with one sequence each, group them by file size - Channel - .fromPath ( fasta ) + fasta .splitFasta ( file:true ) .branch { small: it.size() < split_fasta_cutoff @@ -62,13 +61,16 @@ workflow INPUT_FILTER_SPLIT { .set { fasta_fai } // filter reads - SAMTOOLS_VIEW ( reads, [ [], fasta ], [] ) + fasta + .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } + .set { ch_fasta } + SAMTOOLS_VIEW ( reads, ch_fasta, [] ) ch_versions = ch_versions.mix ( SAMTOOLS_VIEW.out.versions.first() ) // combine reads with splitted references SAMTOOLS_VIEW.out.cram .join ( SAMTOOLS_VIEW.out.crai ) - .map { filtered_reads -> filtered_reads + [interval ?: []] } + .combine(interval) .combine ( fasta_fai ) .set { cram_crai_fasta_fai } diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index da9047e..05776ce 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -18,10 +18,12 @@ workflow INPUT_MERGE { // sort input reads if asked if ( sort_input ) { + SAMTOOLS_SORT( reads ) ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions ) sorted_reads = SAMTOOLS_SORT.out.bam } else { + sorted_reads = reads } @@ -47,10 +49,19 @@ workflow INPUT_MERGE { .set { merged_reads_with_meta } // call samtool merge + fasta + .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } + .set { ch_fasta } + fai + .map { fai -> [ [ 'id': fai.baseName ], fai ] } + .set { ch_fai } + gzi + .map { gzi -> [ [ 'id': gzi.baseName ], gzi ] } + .set { ch_gzi } SAMTOOLS_MERGE( merged_reads_with_meta, - [ [], fasta ], - [ [], fai ], - [ [], gzi ] + ch_fasta, + ch_fai, + ch_gzi ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions ) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 3b36a99..8a933b3 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -14,22 +14,15 @@ def checkPathParamList = [ params.input, params.fasta, params.fai, params.gzi, p for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } // Check mandatory parameters -if (params.input) { input_file = file(params.input) } else { exit 1, 'Input samplesheet not specified!' } -if (params.fasta) { fasta_file = file(params.fasta) } else { exit 1, 'Reference fasta not specified!' } -if (params.fai) { fai_file = file(params.fai) } else { exit 1, 'Reference fasta index not specified!' } - -// Check gzi being given if compressed fasta is provided -if (params.gzi) { - gzi_file = file(params.gzi) -} else if ( params.fasta.endsWith('fasta.gz') ) { - exit 1, 'Reference fasta index gzi file not specified for fasta.gz file!' -} else { - gzi_file = null -} +if (params.input) { ch_input = Channel.fromPath(params.input) } else { exit 1, 'Input samplesheet not specified!' } +if (params.fasta) { ch_fasta = Channel.fromPath(params.fasta) } else { exit 1, 'Reference fasta not specified!' } // Check optional parameters +if (params.fai) { ch_fai = Channel.fromPath(params.fai) } else { ch_fai = Channel.empty() } +if (params.gzi) { ch_gzi = Channel.fromPath(params.gzi) } else { ch_gzi = Channel.empty() } +if (params.interval){ ch_interval = Channel.fromPath(params.interval) } else { ch_interval = Channel.empty() } + if (params.sort_input) { sort_input = params.sort_input } else { sort_input = false } -if (params.interval) { interval_file = file(params.interval) } else { interval_file = null } if (params.split_fasta_cutoff ) { split_fasta_cutoff = params.split_fasta_cutoff } else { split_fasta_cutoff = 100000 } /* @@ -62,6 +55,7 @@ include { DEEPVARIANT_CALLER } from '../subworkflows/local/deepvariant_caller' // MODULE: Installed directly from nf-core/modules // include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' +include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -75,11 +69,33 @@ workflow VARIANTCALLING { ch_versions = Channel.empty() + // + // check reference fasta index given or not + // + if( params.fai == null || ( params.fasta.endsWith('fasta.gz') && params.gzi == null ) ){ + + ch_fasta + .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } + .set { ch_genome } + + SAMTOOLS_FAIDX ( ch_genome, [[], []]) + ch_versions = ch_versions.mix( SAMTOOLS_FAIDX.out.versions ) + + SAMTOOLS_FAIDX.out.fai + .map{ mata, fai -> fai } + .set{ ch_fai } + + SAMTOOLS_FAIDX.out.gzi + .map{ meta, gzi -> gzi } + .set{ ch_gzi } + + } + // // SUBWORKFLOW: Read in samplesheet, validate and stage input files // INPUT_CHECK ( - input_file + ch_input ) ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) @@ -87,9 +103,9 @@ workflow VARIANTCALLING { // SUBWORKFLOW: merge the input reads by sample name // INPUT_MERGE ( - fasta_file, - fai_file, - gzi_file, + ch_fasta, + ch_fai, + ch_gzi, INPUT_CHECK.out.reads, sort_input ) @@ -100,15 +116,15 @@ workflow VARIANTCALLING { // SUBWORKFLOW: split the input fasta file and filter input reads // INPUT_FILTER_SPLIT ( - fasta_file, - fai_file, - gzi_file, + ch_fasta, + ch_fai, + ch_gzi, INPUT_MERGE.out.indexed_merged_reads, - interval_file, + ch_interval, split_fasta_cutoff ) ch_versions = ch_versions.mix(INPUT_FILTER_SPLIT.out.versions) - + // // SUBWORKFLOW: call deepvariant // From 0265cbf8053998245ab5ae14e3caeb50c8f05e63 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 21:52:31 +0100 Subject: [PATCH 054/123] use the first for the reference fasta channel --- subworkflows/local/input_filter_split.nf | 5 ++-- subworkflows/local/input_merge.nf | 35 ++++++++++-------------- 2 files changed, 16 insertions(+), 24 deletions(-) diff --git a/subworkflows/local/input_filter_split.nf b/subworkflows/local/input_filter_split.nf index 95555d7..0482cf2 100644 --- a/subworkflows/local/input_filter_split.nf +++ b/subworkflows/local/input_filter_split.nf @@ -61,9 +61,8 @@ workflow INPUT_FILTER_SPLIT { .set { fasta_fai } // filter reads - fasta - .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } - .set { ch_fasta } + ch_fasta = fasta.map { fasta -> [ [ 'id': fasta.baseName ], fasta ] }.first() + SAMTOOLS_VIEW ( reads, ch_fasta, [] ) ch_versions = ch_versions.mix ( SAMTOOLS_VIEW.out.versions.first() ) diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 05776ce..4e7efa5 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -18,46 +18,39 @@ workflow INPUT_MERGE { // sort input reads if asked if ( sort_input ) { - SAMTOOLS_SORT( reads ) ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions ) sorted_reads = SAMTOOLS_SORT.out.bam - } else { - + } else { sorted_reads = reads } - + // group input reads file by sample name sorted_reads - .map{ it -> [ it[0].sample, it[1] ] } + .map{ meta, bam_cram -> [ meta.sample, bam_cram ] } .groupTuple() .set{ merged_reads } - + // group input meta data together by sample name as well // use the first meta data for the combined reads reads - .map{ it -> [ it[0].sample, it[0] ] } + .map{ meta, bam_cram -> [ meta.sample, meta ] } .groupTuple() - .map { it -> [it[0], it[1][0]] } + .map { sample, meta_list -> [sample, meta_list[0]] } .join( merged_reads ) - .map { it -> [ - [ id: ( it[2].size() == 1 ) ? it[1].sample : it[1].sample + '_combined', - type: it[1].type + .map { sample, meta, bam_cram_list -> [ + [ id: ( bam_cram_list.size() == 1 ) ? sample : sample + '_combined', + type: meta.type ], - it[2] + bam_cram_list ]} .set { merged_reads_with_meta } // call samtool merge - fasta - .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } - .set { ch_fasta } - fai - .map { fai -> [ [ 'id': fai.baseName ], fai ] } - .set { ch_fai } - gzi - .map { gzi -> [ [ 'id': gzi.baseName ], gzi ] } - .set { ch_gzi } + ch_fasta = fasta.map { fasta -> [ [ 'id': fasta.baseName ], fasta ] }.first() + ch_fai = fai.map { fai -> [ [ 'id': fai.baseName ], fai ] }.first() + ch_gzi = gzi.map { gzi -> [ [ 'id': gzi.baseName ], gzi ] }.first() + SAMTOOLS_MERGE( merged_reads_with_meta, ch_fasta, ch_fai, From d4d38cc08e2100a3a48c0beca5932e834b76017a Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 6 Oct 2023 22:17:10 +0100 Subject: [PATCH 055/123] move write-index flag to the config file --- conf/modules.config | 4 ++++ modules/nf-core/samtools/merge/main.nf | 1 - modules/nf-core/samtools/merge/samtools-merge.diff | 8 -------- 3 files changed, 4 insertions(+), 9 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 6672bfc..0deaa90 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -25,6 +25,10 @@ process { ext.prefix = { "${meta.id}_filtered" } } + withName: '.*:INPUT_MERGE:SAMTOOLS_MERGE' { + ext.args = '--write-index' + } + withName: '.*:DEEPVARIANT_CALLER:DEEPVARIANT' { ext.args = '--model_type=PACBIO' } diff --git a/modules/nf-core/samtools/merge/main.nf b/modules/nf-core/samtools/merge/main.nf index 90ddfbe..2dc4008 100644 --- a/modules/nf-core/samtools/merge/main.nf +++ b/modules/nf-core/samtools/merge/main.nf @@ -33,7 +33,6 @@ process SAMTOOLS_MERGE { samtools \\ merge \\ --threads ${task.cpus-1} \\ - --write-index \\ $args \\ ${reference} \\ ${prefix}.${file_type} \\ diff --git a/modules/nf-core/samtools/merge/samtools-merge.diff b/modules/nf-core/samtools/merge/samtools-merge.diff index a740ce0..afe2536 100644 --- a/modules/nf-core/samtools/merge/samtools-merge.diff +++ b/modules/nf-core/samtools/merge/samtools-merge.diff @@ -40,13 +40,5 @@ Changes in module 'nf-core/samtools/merge' path "versions.yml" , emit: versions -@@ -31,6 +33,7 @@ - samtools \\ - merge \\ - --threads ${task.cpus-1} \\ -+ --write-index \\ - $args \\ - ${reference} \\ - ${prefix}.${file_type} \\ ************************************************************ From ea046b9b3a94661860382e185d53cf8fc79b31a9 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Sat, 7 Oct 2023 11:12:47 +0100 Subject: [PATCH 056/123] make sure work file when no interval file given --- subworkflows/local/input_filter_split.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/input_filter_split.nf b/subworkflows/local/input_filter_split.nf index 0482cf2..2467b68 100644 --- a/subworkflows/local/input_filter_split.nf +++ b/subworkflows/local/input_filter_split.nf @@ -69,7 +69,7 @@ workflow INPUT_FILTER_SPLIT { // combine reads with splitted references SAMTOOLS_VIEW.out.cram .join ( SAMTOOLS_VIEW.out.crai ) - .combine(interval) + .combine(interval.ifEmpty([[]])) .combine ( fasta_fai ) .set { cram_crai_fasta_fai } From 7506e81addf69b1960e521438a1e1cdbe7c81e2f Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Sat, 7 Oct 2023 19:47:51 +0100 Subject: [PATCH 057/123] formating and documents --- README.md | 3 ++- docs/usage.md | 24 ++++++++++++------------ subworkflows/local/input_check.nf | 5 ++++- subworkflows/local/input_merge.nf | 5 +++++ workflows/variantcalling.nf | 2 +- 5 files changed, 24 insertions(+), 15 deletions(-) diff --git a/README.md b/README.md index 62da4b2..ed207d1 100644 --- a/README.md +++ b/README.md @@ -18,11 +18,12 @@ On release, automated continuous integration tests run the pipeline on a full-si ## Pipeline summary -The pipleline takes aligned PacBio sample reads (CRAM/BAM files and their index files) from a CSV file and the reference file in FASTA format, and then uses DeepVariant tool to make variant calling. +The pipeline takes aligned PacBio sample reads (CRAM/BAM files) from a CSV file and the reference file in FASTA format, and then uses DeepVariant tool to make variant calling. Steps involved: - Split fasta file into smaller files, normally one sequence per file unless the sequences are too small. +- Merge input BAM/CRAM files together if they have the same sample names. - Filter out reads using the `-F 0x900` option to only retain the primary alignments. - Run DeepVariant using filtered BAM/CRAM files against each of split fasta files. - Merge all VCF and GVCF files generated by DeepVariant by sample together for each input BAM/CRAM file. diff --git a/docs/usage.md b/docs/usage.md index af4811c..71cc1c7 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -2,11 +2,11 @@ ## Introduction -The pipleline takes aligned sample reads (CRAM/BAM files and their index files) from a CSV file and a reference file in FASTA format, and then use DeepVariant to call variants. +The pipeline takes aligned sample reads (CRAM/BAM files) from a CSV file and a reference file in FASTA format, and then use DeepVariant to call variants. ## Samplesheet input -You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use the `input` parameter to specify the samplesheet location. It has to be a comma-separated file with at least 4 columns, and a header row as shown in the examples below. +You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use the `input` parameter to specify the samplesheet location. It has to be a comma-separated file with 3 columns, and a header row as shown in the examples below. ```bash --input '[path to samplesheet file]' @@ -17,21 +17,22 @@ You will need to create a samplesheet with information about the samples you wou The `sample` identifiers have to be the same when you have re-sequenced the same sample more than once e.g. to increase sequencing depth. Below is an example for the same sample sequenced across 3 lanes: ```console -sample,datatype,datafile,indexfile -sample1,pacbio,sample1_1.cram,sample1_1.cram.crai -sample1,pacbio,sample1_2.cram,sample1_3.cram.crai -sample1,pacbio,sample1_3.cram,sample1_3.cram.crai +sample,datatype,datafile +sample1,pacbio,sample1_1.cram +sample1,pacbio,sample1_2.cram +sample1,pacbio,sample1_3.cram ``` +If the given BAM/CRAM files are not sorted, you need to add `--sort_input` in the run command to sort them before merging the files together from the same samples. ### Full samplesheet A final samplesheet file consisting of both BAM or CRAM will look like this. Currently this pipeline only supports Pacbio aligned data. ```console -sample,datatype,datafile,indexfile -sample1,pacbio,/path/to/data/file/file1.bam,/path/to/index/file/file1.bam.bai -sample2,pacbio,/path/to/data/file/file2.cram,/path/to/index/file/file2.cram.crai -sample3,pacbio,/path/to/data/file/file3.bam,/path/to/index/file/file3.bam.csi +sample,datatype,datafile +sample1,pacbio,/path/to/data/file/file1.bam +sample2,pacbio,/path/to/data/file/file2.cram +sample3,pacbio,/path/to/data/file/file3.bam ``` | Column | Description | @@ -39,7 +40,6 @@ sample3,pacbio,/path/to/data/file/file3.bam,/path/to/index/file/file3.bam.csi | `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). | | `datatype` | Sequencing data type. Must be `pacbio`. | | `datafile` | The location for either BAM or CRAM file. | -| `indexfile` | The location for BAM or CRAM index file – BAI, CSI or CRAI. | An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline. @@ -62,7 +62,7 @@ work # Directory containing the nextflow working files # Other nextflow hidden files, eg. history of pipeline runs and old logs. ``` -The pipeline will split the intput fasta file into smaller files to run DeepVariant parallel. You can set the minimum split fasta file size from the command line. For example to set the minimum size as 10K using `--split_fasta_cutoff 10000`. +The pipeline will split the input fasta file into smaller files to run DeepVariant parallel. You can set the minimum split fasta file size from the command line. For example to set the minimum size as 10K using `--split_fasta_cutoff 10000`. ### Updating the pipeline diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index aa0bbc9..c3e0e49 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -12,7 +12,10 @@ workflow INPUT_CHECK { SAMPLESHEET_CHECK ( samplesheet ) .csv .splitCsv ( header:true, sep:',' ) - .map { [[id: it.sample, sample: it.sample.replaceAll(/_T\d+$/, ''), type: it.datatype], file(it.datafile)] } + .map { [ + [ id: it.sample, sample: it.sample.replaceAll(/_T\d+$/, ''), type: it.datatype ], + file(it.datafile) + ] } .set { reads } emit: diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 4e7efa5..58395d0 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -18,11 +18,15 @@ workflow INPUT_MERGE { // sort input reads if asked if ( sort_input ) { + SAMTOOLS_SORT( reads ) ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions ) sorted_reads = SAMTOOLS_SORT.out.bam + } else { + sorted_reads = reads + } // group input reads file by sample name @@ -58,6 +62,7 @@ workflow INPUT_MERGE { ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions ) + // concat merged bam or cram together along with their index file SAMTOOLS_MERGE.out.bam .join(SAMTOOLS_MERGE.out.csi) .concat( diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 8a933b3..a4cf47c 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -78,7 +78,7 @@ workflow VARIANTCALLING { .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } .set { ch_genome } - SAMTOOLS_FAIDX ( ch_genome, [[], []]) + SAMTOOLS_FAIDX ( ch_genome, [[], []] ) ch_versions = ch_versions.mix( SAMTOOLS_FAIDX.out.versions ) SAMTOOLS_FAIDX.out.fai From 4dacfa801896b36c07377bd6623729008796ae94 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Sat, 7 Oct 2023 18:51:06 +0000 Subject: [PATCH 058/123] [automated] Fix linting with Prettier --- docs/usage.md | 11 ++++++----- 1 file changed, 6 insertions(+), 5 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 71cc1c7..1764889 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -22,6 +22,7 @@ sample1,pacbio,sample1_1.cram sample1,pacbio,sample1_2.cram sample1,pacbio,sample1_3.cram ``` + If the given BAM/CRAM files are not sorted, you need to add `--sort_input` in the run command to sort them before merging the files together from the same samples. ### Full samplesheet @@ -35,11 +36,11 @@ sample2,pacbio,/path/to/data/file/file2.cram sample3,pacbio,/path/to/data/file/file3.bam ``` -| Column | Description | -| ----------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | -| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). | -| `datatype` | Sequencing data type. Must be `pacbio`. | -| `datafile` | The location for either BAM or CRAM file. | +| Column | Description | +| ---------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). | +| `datatype` | Sequencing data type. Must be `pacbio`. | +| `datafile` | The location for either BAM or CRAM file. | An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline. From ebaa7e47361d9129a103d75e7c01709de92413c4 Mon Sep 17 00:00:00 2001 From: Guoying Qi <729395+gq1@users.noreply.github.com> Date: Fri, 13 Oct 2023 09:49:21 +0100 Subject: [PATCH 059/123] Update conf/test.config Co-authored-by: Matthieu Muffato --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index f89e185..ad1e731 100644 --- a/conf/test.config +++ b/conf/test.config @@ -25,7 +25,7 @@ params { // Fasta references fasta = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz' - // Reference index file + // Reference index file (optional) // fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.fai' // gzi = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' From 1b2c45e77be1c11cfadfcb2ba608bed08d575651 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 27 Oct 2023 15:50:37 +0100 Subject: [PATCH 060/123] nf-core modules update samtools/merge with conda environment file and maintainers list --- modules.json | 2 +- modules/nf-core/samtools/merge/environment.yml | 6 ++++++ modules/nf-core/samtools/merge/main.nf | 2 +- modules/nf-core/samtools/merge/meta.yml | 6 ++++++ 4 files changed, 14 insertions(+), 2 deletions(-) create mode 100644 modules/nf-core/samtools/merge/environment.yml diff --git a/modules.json b/modules.json index 183f5b3..9781734 100644 --- a/modules.json +++ b/modules.json @@ -32,7 +32,7 @@ }, "samtools/merge": { "branch": "master", - "git_sha": "0460d316170f75f323111b4a2c0a2989f0c32013", + "git_sha": "516189e968feb4ebdd9921806988b4c12b4ac2dc", "installed_by": ["modules"], "patch": "modules/nf-core/samtools/merge/samtools-merge.diff" }, diff --git a/modules/nf-core/samtools/merge/environment.yml b/modules/nf-core/samtools/merge/environment.yml new file mode 100644 index 0000000..04c82f1 --- /dev/null +++ b/modules/nf-core/samtools/merge/environment.yml @@ -0,0 +1,6 @@ +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/merge/main.nf b/modules/nf-core/samtools/merge/main.nf index 2dc4008..0affdbf 100644 --- a/modules/nf-core/samtools/merge/main.nf +++ b/modules/nf-core/samtools/merge/main.nf @@ -2,7 +2,7 @@ process SAMTOOLS_MERGE { tag "$meta.id" label 'process_low' - conda "bioconda::samtools=1.17" + conda 'modules/nf-core/samtools/merge/environment.yml' container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : 'biocontainers/samtools:1.17--h00cdaf9_0' }" diff --git a/modules/nf-core/samtools/merge/meta.yml b/modules/nf-core/samtools/merge/meta.yml index 056a95f..bf0da8b 100644 --- a/modules/nf-core/samtools/merge/meta.yml +++ b/modules/nf-core/samtools/merge/meta.yml @@ -79,3 +79,9 @@ authors: - "@maxulysse" - "@FriederikeHanssen" - "@ramprasadn" +maintainers: + - "@drpatelh" + - "@yuukiiwa " + - "@maxulysse" + - "@FriederikeHanssen" + - "@ramprasadn" From a76298fb06809c6202ebe45ddac6f2ca9b408186 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 1 Nov 2023 19:22:10 +0000 Subject: [PATCH 061/123] Update samtools/merge module and remove its patch. Combine gzi and fai parameters as one. --- conf/test.config | 2 +- modules.json | 2 +- modules/nf-core/samtools/merge/main.nf | 3 +- modules/nf-core/samtools/merge/meta.yml | 4 -- .../samtools/merge/samtools-merge.diff | 44 ------------------- nextflow.config | 1 - nextflow_schema.json | 8 +--- subworkflows/local/input_filter_split.nf | 2 - subworkflows/local/input_merge.nf | 7 +-- workflows/variantcalling.nf | 34 +++++++++----- 10 files changed, 31 insertions(+), 76 deletions(-) delete mode 100644 modules/nf-core/samtools/merge/samtools-merge.diff diff --git a/conf/test.config b/conf/test.config index ad1e731..49b740b 100644 --- a/conf/test.config +++ b/conf/test.config @@ -27,7 +27,7 @@ params { // Reference index file (optional) // fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.fai' - // gzi = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' + fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' // Interval bed file interval = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bed' diff --git a/modules.json b/modules.json index 9781734..956cd97 100644 --- a/modules.json +++ b/modules.json @@ -32,7 +32,7 @@ }, "samtools/merge": { "branch": "master", - "git_sha": "516189e968feb4ebdd9921806988b4c12b4ac2dc", + "git_sha": "e7ce60acc8a33fa17429e966364657a63016e870", "installed_by": ["modules"], "patch": "modules/nf-core/samtools/merge/samtools-merge.diff" }, diff --git a/modules/nf-core/samtools/merge/main.nf b/modules/nf-core/samtools/merge/main.nf index 0affdbf..21f785c 100644 --- a/modules/nf-core/samtools/merge/main.nf +++ b/modules/nf-core/samtools/merge/main.nf @@ -2,7 +2,7 @@ process SAMTOOLS_MERGE { tag "$meta.id" label 'process_low' - conda 'modules/nf-core/samtools/merge/environment.yml' + conda "${moduleDir}/environment.yml" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : 'biocontainers/samtools:1.17--h00cdaf9_0' }" @@ -11,7 +11,6 @@ process SAMTOOLS_MERGE { tuple val(meta), path(input_files, stageAs: "?/*") tuple val(meta2), path(fasta) tuple val(meta3), path(fai) - tuple val(meta4), path(gzi) output: tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam diff --git a/modules/nf-core/samtools/merge/meta.yml b/modules/nf-core/samtools/merge/meta.yml index bf0da8b..2e8f3db 100644 --- a/modules/nf-core/samtools/merge/meta.yml +++ b/modules/nf-core/samtools/merge/meta.yml @@ -43,10 +43,6 @@ input: type: file description: Index of the reference file the CRAM was created with (optional) pattern: "*.fai" - - gzi: - type: file - description: Index of the reference file the CRAM was created with (optional) - pattern: "*.gzi" output: - meta: type: map diff --git a/modules/nf-core/samtools/merge/samtools-merge.diff b/modules/nf-core/samtools/merge/samtools-merge.diff deleted file mode 100644 index afe2536..0000000 --- a/modules/nf-core/samtools/merge/samtools-merge.diff +++ /dev/null @@ -1,44 +0,0 @@ -Changes in module 'nf-core/samtools/merge' ---- modules/nf-core/samtools/merge/meta.yml -+++ modules/nf-core/samtools/merge/meta.yml -@@ -43,6 +43,10 @@ - type: file - description: Index of the reference file the CRAM was created with (optional) - pattern: "*.fai" -+ - gzi: -+ type: file -+ description: Index of the reference file the CRAM was created with (optional) -+ pattern: "*.gzi" - output: - - meta: - type: map -@@ -65,6 +69,10 @@ - type: file - description: BAM index file (optional) - pattern: "*.csi" -+ - crai: -+ type: file -+ description: CRAM index file (optional) -+ pattern: "*.crai" - authors: - - "@drpatelh" - - "@yuukiiwa " - ---- modules/nf-core/samtools/merge/main.nf -+++ modules/nf-core/samtools/merge/main.nf -@@ -11,11 +11,13 @@ - tuple val(meta), path(input_files, stageAs: "?/*") - tuple val(meta2), path(fasta) - tuple val(meta3), path(fai) -+ tuple val(meta4), path(gzi) - - output: - tuple val(meta), path("${prefix}.bam") , optional:true, emit: bam - tuple val(meta), path("${prefix}.cram"), optional:true, emit: cram - tuple val(meta), path("*.csi") , optional:true, emit: csi -+ tuple val(meta), path("*.crai") , optional:true, emit: crai - path "versions.yml" , emit: versions - - - -************************************************************ diff --git a/nextflow.config b/nextflow.config index 4659669..c1e8ac3 100644 --- a/nextflow.config +++ b/nextflow.config @@ -14,7 +14,6 @@ params { sort_input = false fasta = null fai = null - gzi = null interval = null split_fasta_cutoff = 100000 diff --git a/nextflow_schema.json b/nextflow_schema.json index dc6428b..66cfeda 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -49,15 +49,11 @@ "properties": { "fasta": { "type": "string", - "description": "Path to FASTA genome file, either fasta or fast.gz" + "description": "Path to FASTA genome file, either fasta or fast.gz." }, "fai": { "type": "string", - "description": "Path to the index file of the FASTA genome file." - }, - "gzi": { - "type": "string", - "description": "Path to the gzi index file of the FASTA genome file. Required if fasta in gz format." + "description": "Path to the index file of the FASTA genome file, either fai or gzi." }, "interval": { "type": "string", diff --git a/subworkflows/local/input_filter_split.nf b/subworkflows/local/input_filter_split.nf index 2467b68..dc0710f 100644 --- a/subworkflows/local/input_filter_split.nf +++ b/subworkflows/local/input_filter_split.nf @@ -9,8 +9,6 @@ include { CAT_CAT } from '../../modules/nf-core/cat/cat/main' workflow INPUT_FILTER_SPLIT { take: fasta // file: /path/to/genome.fasta or /path/to/genome.fasta.gz - fai // file: /path/to/genome.*.fai - gzi // file: /path/to/genome.fasta.gz.gzi or null reads // [ val(meta), data, index ] interval // file: /path/to/intervals.bed split_fasta_cutoff // val(min_file_size) diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 58395d0..1c8a588 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -8,8 +8,7 @@ include { SAMTOOLS_SORT } from '../../modules/nf-core/samtools/sort' workflow INPUT_MERGE { take: fasta // file: /path/to/genome.fasta or /path/to/genome.fasta.gz - fai // file: /path/to/genome.*.fai - gzi // file: /path/to/genome.fasta.gz.gzi or null + fai // file: /path/to/genome.*.fai or /path/to/genome.fasta.gz.gzi reads // channel: [ val(meta), data ] sort_input // bollean: true or false @@ -53,12 +52,10 @@ workflow INPUT_MERGE { // call samtool merge ch_fasta = fasta.map { fasta -> [ [ 'id': fasta.baseName ], fasta ] }.first() ch_fai = fai.map { fai -> [ [ 'id': fai.baseName ], fai ] }.first() - ch_gzi = gzi.map { gzi -> [ [ 'id': gzi.baseName ], gzi ] }.first() SAMTOOLS_MERGE( merged_reads_with_meta, ch_fasta, - ch_fai, - ch_gzi + ch_fai ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions ) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index a4cf47c..68e2fef 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -10,7 +10,7 @@ def summary_params = NfcoreSchema.paramsSummaryMap(workflow, params) WorkflowVariantcalling.initialise(params, log) // Check input path parameters to see if they exist -def checkPathParamList = [ params.input, params.fasta, params.fai, params.gzi, params.interval ] +def checkPathParamList = [ params.input, params.fasta, params.fai, params.interval ] for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } // Check mandatory parameters @@ -18,8 +18,17 @@ if (params.input) { ch_input = Channel.fromPath(params.input) } else { exit 1, ' if (params.fasta) { ch_fasta = Channel.fromPath(params.fasta) } else { exit 1, 'Reference fasta not specified!' } // Check optional parameters -if (params.fai) { ch_fai = Channel.fromPath(params.fai) } else { ch_fai = Channel.empty() } -if (params.gzi) { ch_gzi = Channel.fromPath(params.gzi) } else { ch_gzi = Channel.empty() } +if (params.fai){ + if( ( params.fasta.endsWith('.gz') && params.fai.endsWith('.fai') ) + || + ( !params.fasta.endsWith('.gz') && params.fai.endsWith('.gzi') ) + ){ + exit 1, 'Reference fasta and its index file format not matched!' + } + ch_fai = Channel.fromPath(params.fai) +} else { + ch_fai = Channel.empty() +} if (params.interval){ ch_interval = Channel.fromPath(params.interval) } else { ch_interval = Channel.empty() } if (params.sort_input) { sort_input = params.sort_input } else { sort_input = false } @@ -72,7 +81,7 @@ workflow VARIANTCALLING { // // check reference fasta index given or not // - if( params.fai == null || ( params.fasta.endsWith('fasta.gz') && params.gzi == null ) ){ + if( params.fai == null ){ ch_fasta .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } @@ -80,15 +89,23 @@ workflow VARIANTCALLING { SAMTOOLS_FAIDX ( ch_genome, [[], []] ) ch_versions = ch_versions.mix( SAMTOOLS_FAIDX.out.versions ) - + SAMTOOLS_FAIDX.out.fai .map{ mata, fai -> fai } .set{ ch_fai } - + SAMTOOLS_FAIDX.out.gzi .map{ meta, gzi -> gzi } .set{ ch_gzi } + if( params.fasta.endsWith('.gz') ){ + ch_index = ch_gzi + }else{ + ch_index = ch_fai + } + + }else{ + ch_index = ch_fai } // @@ -104,8 +121,7 @@ workflow VARIANTCALLING { // INPUT_MERGE ( ch_fasta, - ch_fai, - ch_gzi, + ch_index, INPUT_CHECK.out.reads, sort_input ) @@ -117,8 +133,6 @@ workflow VARIANTCALLING { // INPUT_FILTER_SPLIT ( ch_fasta, - ch_fai, - ch_gzi, INPUT_MERGE.out.indexed_merged_reads, ch_interval, split_fasta_cutoff From 3cf953d19c3f7c9fc40e5f7878d379364286bd14 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 1 Nov 2023 20:42:24 +0000 Subject: [PATCH 062/123] remove sort_input params. Always sort the input before merging. --- conf/test.config | 2 +- docs/usage.md | 2 -- nextflow.config | 1 - nextflow_schema.json | 4 ---- subworkflows/local/input_merge.nf | 35 +++++++++++++------------------ workflows/variantcalling.nf | 2 -- 6 files changed, 16 insertions(+), 30 deletions(-) diff --git a/conf/test.config b/conf/test.config index 49b740b..01515e9 100644 --- a/conf/test.config +++ b/conf/test.config @@ -27,7 +27,7 @@ params { // Reference index file (optional) // fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.fai' - fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' + // fai = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz.gzi' // Interval bed file interval = 'https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/analysis/icCanRufa1/read_mapping/pacbio/GCA_947369205.1.unmasked.pacbio.icCanRufa1_0_3.bed' diff --git a/docs/usage.md b/docs/usage.md index 1764889..46b74b3 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -23,8 +23,6 @@ sample1,pacbio,sample1_2.cram sample1,pacbio,sample1_3.cram ``` -If the given BAM/CRAM files are not sorted, you need to add `--sort_input` in the run command to sort them before merging the files together from the same samples. - ### Full samplesheet A final samplesheet file consisting of both BAM or CRAM will look like this. Currently this pipeline only supports Pacbio aligned data. diff --git a/nextflow.config b/nextflow.config index c1e8ac3..399b382 100644 --- a/nextflow.config +++ b/nextflow.config @@ -11,7 +11,6 @@ params { // Input options input = null - sort_input = false fasta = null fai = null interval = null diff --git a/nextflow_schema.json b/nextflow_schema.json index 66cfeda..d40c501 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -28,10 +28,6 @@ "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.", "fa_icon": "fas fa-folder-open" }, - "sort_input": { - "type": "boolean", - "description": "Boolean whether to sort input reads files" - }, "email": { "type": "string", "description": "Email address for completion summary.", diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 1c8a588..90bb82f 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -10,50 +10,45 @@ workflow INPUT_MERGE { fasta // file: /path/to/genome.fasta or /path/to/genome.fasta.gz fai // file: /path/to/genome.*.fai or /path/to/genome.fasta.gz.gzi reads // channel: [ val(meta), data ] - sort_input // bollean: true or false main: ch_versions = Channel.empty() - - // sort input reads if asked - if ( sort_input ) { - - SAMTOOLS_SORT( reads ) - ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions ) - sorted_reads = SAMTOOLS_SORT.out.bam - } else { - - sorted_reads = reads + // group input meta data together by sample name + reads + .map{ meta, bam_cram -> [ meta.sample, meta ] } + .groupTuple() + .set{ grouped_reads_meta } - } + // sort input reads + SAMTOOLS_SORT( reads ) + ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions ) + sorted_reads = SAMTOOLS_SORT.out.bam // group input reads file by sample name sorted_reads .map{ meta, bam_cram -> [ meta.sample, bam_cram ] } .groupTuple() - .set{ merged_reads } + .set{ grouped_reads } - // group input meta data together by sample name as well + // join grouped reads and meta // use the first meta data for the combined reads - reads - .map{ meta, bam_cram -> [ meta.sample, meta ] } - .groupTuple() + grouped_reads_meta .map { sample, meta_list -> [sample, meta_list[0]] } - .join( merged_reads ) + .join( grouped_reads ) .map { sample, meta, bam_cram_list -> [ [ id: ( bam_cram_list.size() == 1 ) ? sample : sample + '_combined', type: meta.type ], bam_cram_list ]} - .set { merged_reads_with_meta } + .set { grouped_reads_with_meta } // call samtool merge ch_fasta = fasta.map { fasta -> [ [ 'id': fasta.baseName ], fasta ] }.first() ch_fai = fai.map { fai -> [ [ 'id': fai.baseName ], fai ] }.first() - SAMTOOLS_MERGE( merged_reads_with_meta, + SAMTOOLS_MERGE( grouped_reads_with_meta, ch_fasta, ch_fai ) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 68e2fef..99c35e5 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -31,7 +31,6 @@ if (params.fai){ } if (params.interval){ ch_interval = Channel.fromPath(params.interval) } else { ch_interval = Channel.empty() } -if (params.sort_input) { sort_input = params.sort_input } else { sort_input = false } if (params.split_fasta_cutoff ) { split_fasta_cutoff = params.split_fasta_cutoff } else { split_fasta_cutoff = 100000 } /* @@ -123,7 +122,6 @@ workflow VARIANTCALLING { ch_fasta, ch_index, INPUT_CHECK.out.reads, - sort_input ) ch_versions = ch_versions.mix(INPUT_MERGE.out.versions) From d6fa00f37eb2729b6c67b3d0a4d36f02e1536b35 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 2 Nov 2023 11:41:59 +0000 Subject: [PATCH 063/123] only validate the sample sheet not transform the sample names --- bin/check_samplesheet.py | 22 +++++++--------------- subworkflows/local/input_check.nf | 2 +- 2 files changed, 8 insertions(+), 16 deletions(-) diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py index 52af146..d088e65 100755 --- a/bin/check_samplesheet.py +++ b/bin/check_samplesheet.py @@ -1,6 +1,6 @@ #!/usr/bin/env python -"""Provide a command line tool to validate and transform tabular samplesheets.""" +"""Provide a command line tool to validate tabular samplesheets.""" import argparse @@ -55,9 +55,9 @@ def __init__( self._type_col = type_col self._file_col = file_col self._seen = set() - self.modified = [] + self.validated = [] - def validate_and_transform(self, row): + def validate(self, row): """ Perform all validations on the given row. @@ -70,7 +70,7 @@ def validate_and_transform(self, row): self._validate_type(row) self._validate_data_file(row) self._seen.add((row[self._sample_col], row[self._file_col])) - self.modified.append(row) + self.validated.append(row) def _validate_sample(self, row): """Assert that the sample name exists and convert spaces to underscores.""" @@ -105,17 +105,9 @@ def validate_unique_samples(self): """ Assert that the combination of sample name and data filename is unique. - In addition to the validation, also rename all samples to have a suffix of _T{n}, where n is the - number of times the same sample exist, but with different files, e.g., multiple runs per experiment. - """ - if len(self._seen) != len(self.modified): + if len(self._seen) != len(self.validated): raise AssertionError("The combination of sample name and data file must be unique.") - seen = Counter() - for row in self.modified: - sample = row[self._sample_col] - seen[sample] += 1 - row[self._sample_col] = f"{sample}_T{seen[sample]}" def read_head(handle, num_lines=10): @@ -195,7 +187,7 @@ def check_samplesheet(file_in, file_out): checker = RowChecker() for i, row in enumerate(reader): try: - checker.validate_and_transform(row) + checker.validate(row) except AssertionError as error: logger.critical(f"{str(error)} On line {i + 2}.") sys.exit(1) @@ -205,7 +197,7 @@ def check_samplesheet(file_in, file_out): with file_out.open(mode="w", newline="") as out_handle: writer = csv.DictWriter(out_handle, header, delimiter=",") writer.writeheader() - for row in checker.modified: + for row in checker.validated: writer.writerow(row) diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index c3e0e49..7e9f667 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -13,7 +13,7 @@ workflow INPUT_CHECK { .csv .splitCsv ( header:true, sep:',' ) .map { [ - [ id: it.sample, sample: it.sample.replaceAll(/_T\d+$/, ''), type: it.datatype ], + [ id: it.sample, sample: it.sample, type: it.datatype ], file(it.datafile) ] } .set { reads } From 3e8705cdee3e4adb71309ab4c40ba246c3cf9a73 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 2 Nov 2023 11:42:32 +0000 Subject: [PATCH 064/123] update fai file for the full test and formating --- conf/test_full.config | 3 +-- workflows/variantcalling.nf | 5 +++-- 2 files changed, 4 insertions(+), 4 deletions(-) diff --git a/conf/test_full.config b/conf/test_full.config index 8532e0d..3a2d38e 100644 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -23,6 +23,5 @@ params { fasta = '/lustre/scratch124/tol/projects/darwin/data/insects/Cantharis_rufa/assembly/release/icCanRufa1.1/insdc/GCA_947369205.1.fasta.gz' // Reference index file - fai = '/lustre/scratch124/tol/projects/darwin/data/insects/Cantharis_rufa/assembly/release/icCanRufa1.1/insdc/GCA_947369205.1.fasta.gz.fai' - gzi = '/lustre/scratch124/tol/projects/darwin/data/insects/Cantharis_rufa/assembly/release/icCanRufa1.1/insdc/GCA_947369205.1.fasta.gz.gzi' + fai = '/lustre/scratch124/tol/projects/darwin/data/insects/Cantharis_rufa/assembly/release/icCanRufa1.1/insdc/GCA_947369205.1.fasta.gz.gzi' } diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 99c35e5..82267f7 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -25,10 +25,11 @@ if (params.fai){ ){ exit 1, 'Reference fasta and its index file format not matched!' } - ch_fai = Channel.fromPath(params.fai) + ch_fai = Channel.fromPath(params.fai) } else { - ch_fai = Channel.empty() + ch_fai = Channel.empty() } + if (params.interval){ ch_interval = Channel.fromPath(params.interval) } else { ch_interval = Channel.empty() } if (params.split_fasta_cutoff ) { split_fasta_cutoff = params.split_fasta_cutoff } else { split_fasta_cutoff = 100000 } From 2c10d8db2e054ce77ea6256472be5ae609e6827f Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 6 Nov 2023 15:18:18 +0000 Subject: [PATCH 065/123] nf-core modules install vcftools --- modules.json | 5 + modules/nf-core/vcftools/environment.yml | 6 + modules/nf-core/vcftools/main.nf | 123 ++++++++++ modules/nf-core/vcftools/meta.yml | 292 +++++++++++++++++++++++ 4 files changed, 426 insertions(+) create mode 100644 modules/nf-core/vcftools/environment.yml create mode 100644 modules/nf-core/vcftools/main.nf create mode 100644 modules/nf-core/vcftools/meta.yml diff --git a/modules.json b/modules.json index 956cd97..8fa7e99 100644 --- a/modules.json +++ b/modules.json @@ -45,6 +45,11 @@ "branch": "master", "git_sha": "3ffae3598260a99e8db3207dead9f73f87f90d1f", "installed_by": ["modules"] + }, + "vcftools": { + "branch": "master", + "git_sha": "8fc1d24c710ebe1d5de0f2447ec9439fd3d9d66a", + "installed_by": ["modules"] } } } diff --git a/modules/nf-core/vcftools/environment.yml b/modules/nf-core/vcftools/environment.yml new file mode 100644 index 0000000..875817e --- /dev/null +++ b/modules/nf-core/vcftools/environment.yml @@ -0,0 +1,6 @@ +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::vcftools=0.1.16 diff --git a/modules/nf-core/vcftools/main.nf b/modules/nf-core/vcftools/main.nf new file mode 100644 index 0000000..0153a60 --- /dev/null +++ b/modules/nf-core/vcftools/main.nf @@ -0,0 +1,123 @@ +process VCFTOOLS { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/vcftools:0.1.16--he513fc3_4' : + 'biocontainers/vcftools:0.1.16--he513fc3_4' }" + + input: + // Owing to the nature of vcftools we here provide solutions to working with optional bed files and optional + // alternative variant files, for use with the 'diff' suite of tools. + // Other optional input files can be utilised in a similar way to below but we do not exhaustively itterate through all + // possible options. Instead we leave that to the user. + tuple val(meta), path(variant_file) + path bed + path diff_variant_file + + output: + tuple val(meta), path("*.vcf") , optional:true, emit: vcf + tuple val(meta), path("*.bcf") , optional:true, emit: bcf + tuple val(meta), path("*.frq") , optional:true, emit: frq + tuple val(meta), path("*.frq.count") , optional:true, emit: frq_count + tuple val(meta), path("*.idepth") , optional:true, emit: idepth + tuple val(meta), path("*.ldepth") , optional:true, emit: ldepth + tuple val(meta), path("*.ldepth.mean") , optional:true, emit: ldepth_mean + tuple val(meta), path("*.gdepth") , optional:true, emit: gdepth + tuple val(meta), path("*.hap.ld") , optional:true, emit: hap_ld + tuple val(meta), path("*.geno.ld") , optional:true, emit: geno_ld + tuple val(meta), path("*.geno.chisq") , optional:true, emit: geno_chisq + tuple val(meta), path("*.list.hap.ld") , optional:true, emit: list_hap_ld + tuple val(meta), path("*.list.geno.ld") , optional:true, emit: list_geno_ld + tuple val(meta), path("*.interchrom.hap.ld") , optional:true, emit: interchrom_hap_ld + tuple val(meta), path("*.interchrom.geno.ld") , optional:true, emit: interchrom_geno_ld + tuple val(meta), path("*.TsTv") , optional:true, emit: tstv + tuple val(meta), path("*.TsTv.summary") , optional:true, emit: tstv_summary + tuple val(meta), path("*.TsTv.count") , optional:true, emit: tstv_count + tuple val(meta), path("*.TsTv.qual") , optional:true, emit: tstv_qual + tuple val(meta), path("*.FILTER.summary") , optional:true, emit: filter_summary + tuple val(meta), path("*.sites.pi") , optional:true, emit: sites_pi + tuple val(meta), path("*.windowed.pi") , optional:true, emit: windowed_pi + tuple val(meta), path("*.weir.fst") , optional:true, emit: weir_fst + tuple val(meta), path("*.het") , optional:true, emit: heterozygosity + tuple val(meta), path("*.hwe") , optional:true, emit: hwe + tuple val(meta), path("*.Tajima.D") , optional:true, emit: tajima_d + tuple val(meta), path("*.ifreqburden") , optional:true, emit: freq_burden + tuple val(meta), path("*.LROH") , optional:true, emit: lroh + tuple val(meta), path("*.relatedness") , optional:true, emit: relatedness + tuple val(meta), path("*.relatedness2") , optional:true, emit: relatedness2 + tuple val(meta), path("*.lqual") , optional:true, emit: lqual + tuple val(meta), path("*.imiss") , optional:true, emit: missing_individual + tuple val(meta), path("*.lmiss") , optional:true, emit: missing_site + tuple val(meta), path("*.snpden") , optional:true, emit: snp_density + tuple val(meta), path("*.kept.sites") , optional:true, emit: kept_sites + tuple val(meta), path("*.removed.sites") , optional:true, emit: removed_sites + tuple val(meta), path("*.singletons") , optional:true, emit: singeltons + tuple val(meta), path("*.indel.hist") , optional:true, emit: indel_hist + tuple val(meta), path("*.hapcount") , optional:true, emit: hapcount + tuple val(meta), path("*.mendel") , optional:true, emit: mendel + tuple val(meta), path("*.FORMAT") , optional:true, emit: format + tuple val(meta), path("*.INFO") , optional:true, emit: info + tuple val(meta), path("*.012") , optional:true, emit: genotypes_matrix + tuple val(meta), path("*.012.indv") , optional:true, emit: genotypes_matrix_individual + tuple val(meta), path("*.012.pos") , optional:true, emit: genotypes_matrix_position + tuple val(meta), path("*.impute.hap") , optional:true, emit: impute_hap + tuple val(meta), path("*.impute.hap.legend") , optional:true, emit: impute_hap_legend + tuple val(meta), path("*.impute.hap.indv") , optional:true, emit: impute_hap_indv + tuple val(meta), path("*.ldhat.sites") , optional:true, emit: ldhat_sites + tuple val(meta), path("*.ldhat.locs") , optional:true, emit: ldhat_locs + tuple val(meta), path("*.BEAGLE.GL") , optional:true, emit: beagle_gl + tuple val(meta), path("*.BEAGLE.PL") , optional:true, emit: beagle_pl + tuple val(meta), path("*.ped") , optional:true, emit: ped + tuple val(meta), path("*.map") , optional:true, emit: map_ + tuple val(meta), path("*.tped") , optional:true, emit: tped + tuple val(meta), path("*.tfam") , optional:true, emit: tfam + tuple val(meta), path("*.diff.sites_in_files") , optional:true, emit: diff_sites_in_files + tuple val(meta), path("*.diff.indv_in_files") , optional:true, emit: diff_indv_in_files + tuple val(meta), path("*.diff.sites") , optional:true, emit: diff_sites + tuple val(meta), path("*.diff.indv") , optional:true, emit: diff_indv + tuple val(meta), path("*.diff.discordance.matrix"), optional:true, emit: diff_discd_matrix + tuple val(meta), path("*.diff.switch") , optional:true, emit: diff_switch_error + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def args_list = args.tokenize() + + def bed_arg = (args.contains('--bed')) ? "--bed ${bed}" : + (args.contains('--exclude-bed')) ? "--exclude-bed ${bed}" : + (args.contains('--hapcount')) ? "--hapcount ${bed}" : '' + args_list.removeIf { it.contains('--bed') } + args_list.removeIf { it.contains('--exclude-bed') } + args_list.removeIf { it.contains('--hapcount') } + + def diff_variant_arg = (args.contains('--diff')) ? "--diff ${diff_variant_file}" : + (args.contains('--gzdiff')) ? "--gzdiff ${diff_variant_file}" : + (args.contains('--diff-bcf')) ? "--diff-bcf ${diff_variant_file}" : '' + args_list.removeIf { it.contains('--diff') } + args_list.removeIf { it.contains('--gzdiff') } + args_list.removeIf { it.contains('--diff-bcf') } + + def input_file = ("$variant_file".endsWith(".vcf")) ? "--vcf ${variant_file}" : + ("$variant_file".endsWith(".vcf.gz")) ? "--gzvcf ${variant_file}" : + ("$variant_file".endsWith(".bcf")) ? "--bcf ${variant_file}" : '' + + """ + vcftools \\ + $input_file \\ + --out $prefix \\ + ${args_list.join(' ')} \\ + $bed_arg \\ + $diff_variant_arg + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + vcftools: \$(echo \$(vcftools --version 2>&1) | sed 's/^.*VCFtools (//;s/).*//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/vcftools/meta.yml b/modules/nf-core/vcftools/meta.yml new file mode 100644 index 0000000..f361db4 --- /dev/null +++ b/modules/nf-core/vcftools/meta.yml @@ -0,0 +1,292 @@ +name: vcftools +description: A set of tools written in Perl and C++ for working with VCF files +keywords: + - VCF + - sort +tools: + - vcftools: + description: A set of tools written in Perl and C++ for working with VCF files. This package only contains the C++ libraries whereas the package perl-vcftools-vcf contains the perl libraries + homepage: http://vcftools.sourceforge.net/ + documentation: http://vcftools.sourceforge.net/man_latest.html + licence: ["LGPL"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - variant_file: + type: file + description: variant input file which can be vcf, vcf.gz, or bcf format. + - bed: + type: file + description: bed file which can be used with different arguments in vcftools (optional) + - diff_variant_file: + type: file + description: secondary variant file which can be used with the 'diff' suite of tools (optional) +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - vcf: + type: file + description: vcf file (optional) + pattern: "*.vcf" + - bcf: + type: file + description: bcf file (optional) + pattern: "*.bcf" + - frq: + type: file + description: Allele frequency for each site (optional) + pattern: "*.frq" + - frq_count: + type: file + description: Allele counts for each site (optional) + pattern: "*.frq.count" + - idepth: + type: file + description: mean depth per individual (optional) + pattern: "*.idepth" + - ldepth: + type: file + description: depth per site summed across individuals (optional) + pattern: "*.ildepth" + - ldepth_mean: + type: file + description: mean depth per site calculated across individuals (optional) + pattern: "*.ldepth.mean" + - gdepth: + type: file + description: depth for each genotype in vcf file (optional) + pattern: "*.gdepth" + - hap_ld: + type: file + description: r2, D, and D’ statistics using phased haplotypes (optional) + pattern: "*.hap.ld" + - geno_ld: + type: file + description: squared correlation coefficient between genotypes encoded as 0, 1 and 2 to represent the number of non-reference alleles in each individual (optional) + pattern: "*.geno.ld" + - geno_chisq: + type: file + description: test for genotype independence via the chi-squared statistic (optional) + pattern: "*.geno.chisq" + - list_hap_ld: + type: file + description: r2 statistics of the sites contained in the provided input file verses all other sites (optional) + pattern: "*.list.hap.ld" + - list_geno_ld: + type: file + description: r2 statistics of the sites contained in the provided input file verses all other sites (optional) + pattern: "*.list.geno.ld" + - interchrom_hap_ld: + type: file + description: r2 statistics for sites (haplotypes) on different chromosomes (optional) + pattern: "*.interchrom.hap.ld" + - interchrom_geno_ld: + type: file + description: r2 statistics for sites (genotypes) on different chromosomes (optional) + pattern: "*.interchrom.geno.ld" + - tstv: + type: file + description: Transition / Transversion ratio in bins of size defined in options (optional) + pattern: "*.TsTv" + - tstv_summary: + type: file + description: Summary of all Transitions and Transversions (optional) + pattern: "*.TsTv.summary" + - tstv_count: + type: file + description: Transition / Transversion ratio as a function of alternative allele count (optional) + pattern: "*.TsTv.count" + - tstv_qual: + type: file + description: Transition / Transversion ratio as a function of SNP quality threshold (optional) + pattern: "*.TsTv.qual" + - filter_summary: + type: file + description: Summary of the number of SNPs and Ts/Tv ratio for each FILTER category (optional) + pattern: "*.FILTER.summary" + - sites_pi: + type: file + description: Nucleotide divergency on a per-site basis (optional) + pattern: "*.sites.pi" + - windowed_pi: + type: file + description: Nucleotide diversity in windows, with window size determined by options (optional) + pattern: "*windowed.pi" + - weir_fst: + type: file + description: Fst estimate from Weir and Cockerham’s 1984 paper (optional) + pattern: "*.weir.fst" + - heterozygosity: + type: file + description: Heterozygosity on a per-individual basis (optional) + pattern: "*.het" + - hwe: + type: file + description: Contains the Observed numbers of Homozygotes and Heterozygotes and the corresponding Expected numbers under HWE (optional) + pattern: "*.hwe" + - tajima_d: + type: file + description: Tajima’s D statistic in bins with size of the specified number in options (optional) + pattern: "*.Tajima.D" + - freq_burden: + type: file + description: Number of variants within each individual of a specific frequency in options (optional) + pattern: "*.ifreqburden" + - lroh: + type: file + description: Long Runs of Homozygosity (optional) + pattern: "*.LROH" + - relatedness: + type: file + description: Relatedness statistic based on the method of Yang et al, Nature Genetics 2010 (doi:10.1038/ng.608) (optional) + pattern: "*.relatedness" + - relatedness2: + type: file + description: Relatedness statistic based on the method of Manichaikul et al., BIOINFORMATICS 2010 (doi:10.1093/bioinformatics/btq559) (optional) + pattern: "*.relatedness2" + - lqual: + type: file + description: per-site SNP quality (optional) + pattern: "*.lqual" + - missing_individual: + type: file + description: Missingness on a per-individual basis (optional) + pattern: "*.imiss" + - missing_site: + type: file + description: Missingness on a per-site basis (optional) + pattern: "*.lmiss" + - snp_density: + type: file + description: Number and density of SNPs in bins of size defined by option (optional) + pattern: "*.snpden" + - kept_sites: + type: file + description: All sites that have been kept after filtering (optional) + pattern: "*.kept.sites" + - removed_sites: + type: file + description: All sites that have been removed after filtering (optional) + pattern: "*.removed.sites" + - singeltons: + type: file + description: Location of singletons, and the individual they occur in (optional) + pattern: "*.singeltons" + - indel_hist: + type: file + description: Histogram file of the length of all indels (including SNPs) (optional) + pattern: "*.indel_hist" + - hapcount: + type: file + description: Unique haplotypes within user specified bins (optional) + pattern: "*.hapcount" + - mendel: + type: file + description: Mendel errors identified in trios (optional) + pattern: "*.mendel" + - format: + type: file + description: Extracted information from the genotype fields in the VCF file relating to a specfied FORMAT identifier (optional) + pattern: "*.FORMAT" + - info: + type: file + description: Extracted information from the INFO field in the VCF file (optional) + pattern: "*.INFO" + - genotypes_matrix: + type: file + description: | + Genotypes output as large matrix. + Genotypes of each individual on a separate line. + Genotypes are represented as 0, 1 and 2, where the number represent that number of non-reference alleles. + Missing genotypes are represented by -1 (optional) + pattern: "*.012" + - genotypes_matrix_individual: + type: file + description: Details the individuals included in the main genotypes_matrix file (optional) + pattern: "*.012.indv" + - genotypes_matrix_position: + type: file + description: Details the site locations included in the main genotypes_matrix file (optional) + pattern: "*.012.pos" + - impute_hap: + type: file + description: Phased haplotypes in IMPUTE reference-panel format (optional) + pattern: "*.impute.hap" + - impute_hap_legend: + type: file + description: Impute haplotype legend file (optional) + pattern: "*.impute.hap.legend" + - impute_hap_indv: + type: file + description: Impute haplotype individuals file (optional) + pattern: "*.impute.hap.indv" + - ldhat_sites: + type: file + description: Output data in LDhat format, sites (optional) + pattern: "*.ldhat.sites" + - ldhat_locs: + type: file + description: output data in LDhat format, locations (optional) + pattern: "*.ldhat.locs" + - beagle_gl: + type: file + description: Genotype likelihoods for biallelic sites (optional) + pattern: "*.BEAGLE.GL" + - beagle_pl: + type: file + description: Genotype likelihoods for biallelic sites (optional) + pattern: "*.BEAGLE.PL" + - ped: + type: file + description: output the genotype data in PLINK PED format (optional) + pattern: "*.ped" + - map_: + type: file + description: output the genotype data in PLINK PED format (optional) + pattern: "*.map" + - tped: + type: file + description: output the genotype data in PLINK PED format (optional) + pattern: "*.tped" + - tfam: + type: file + description: output the genotype data in PLINK PED format (optional) + pattern: "*.tfam" + - diff_sites_in_files: + type: file + description: Sites that are common / unique to each file specified in optional inputs (optional) + pattern: "*.diff.sites.in.files" + - diff_indv_in_files: + type: file + description: Individuals that are common / unique to each file specified in optional inputs (optional) + pattern: "*.diff.indv.in.files" + - diff_sites: + type: file + description: Discordance on a site by site basis, specified in optional inputs (optional) + pattern: "*.diff.sites" + - diff_indv: + type: file + description: Discordance on a individual by individual basis, specified in optional inputs (optional) + pattern: "*.diff.indv" + - diff_discd_matrix: + type: file + description: Discordance matrix between files specified in optional inputs (optional) + pattern: "*.diff.discordance.matrix" + - diff_switch_error: + type: file + description: Switch errors found between sites (optional) + pattern: "*.diff.switch" +authors: + - "@Mark-S-Hill" +maintainers: + - "@Mark-S-Hill" From 124c5850f3934902092d1776b78a7705aebe339c Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 6 Nov 2023 15:49:06 +0000 Subject: [PATCH 066/123] nf-core modules update deepvariant --- modules.json | 2 +- modules/nf-core/deepvariant/environment.yml | 4 ++++ modules/nf-core/deepvariant/meta.yml | 6 +++--- 3 files changed, 8 insertions(+), 4 deletions(-) create mode 100644 modules/nf-core/deepvariant/environment.yml diff --git a/modules.json b/modules.json index 8fa7e99..3ce8a13 100644 --- a/modules.json +++ b/modules.json @@ -22,7 +22,7 @@ }, "deepvariant": { "branch": "master", - "git_sha": "ed67f2fadd6d2a155b296f728e6b1f8c92ddc1a6", + "git_sha": "516189e968feb4ebdd9921806988b4c12b4ac2dc", "installed_by": ["modules"] }, "samtools/faidx": { diff --git a/modules/nf-core/deepvariant/environment.yml b/modules/nf-core/deepvariant/environment.yml new file mode 100644 index 0000000..bcd3a8b --- /dev/null +++ b/modules/nf-core/deepvariant/environment.yml @@ -0,0 +1,4 @@ +channels: + - conda-forge + - bioconda + - defaults diff --git a/modules/nf-core/deepvariant/meta.yml b/modules/nf-core/deepvariant/meta.yml index c7d11ae..a50dc57 100644 --- a/modules/nf-core/deepvariant/meta.yml +++ b/modules/nf-core/deepvariant/meta.yml @@ -12,7 +12,6 @@ tools: tool_dev_url: https://github.com/google/deepvariant doi: "10.1038/nbt.4235" licence: ["BSD-3-clause"] - input: - meta: type: map @@ -58,7 +57,6 @@ input: type: file description: GZI index of reference fasta file pattern: "*.gzi" - output: - meta: type: map @@ -77,7 +75,9 @@ output: type: file description: File containing software version pattern: "*.{version.txt}" - authors: - "@abhi18av" - "@ramprasadn" +maintainers: + - "@abhi18av" + - "@ramprasadn" From 908e9f43e3543bfefd837214f2cef11597905c71 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 7 Nov 2023 16:41:59 +0000 Subject: [PATCH 067/123] create a sub workflow to call vcftools to calculate per site nucleotide diversity and heterozygosity --- conf/modules.config | 18 +++++++++++++++++ subworkflows/local/process_vcf.nf | 32 +++++++++++++++++++++++++++++++ workflows/variantcalling.nf | 26 +++++++++++++++++++++---- 3 files changed, 72 insertions(+), 4 deletions(-) create mode 100644 subworkflows/local/process_vcf.nf diff --git a/conf/modules.config b/conf/modules.config index 0deaa90..660809c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -51,6 +51,24 @@ process { ] } + withName: '.*:PROCESS_VCF:VCFTOOLS_SITE_PI' { + ext.args = '--site-pi' + publishDir = [ + path: { "${params.outdir}/variant_calling" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + + withName: '.*:PROCESS_VCF:VCFTOOLS_HET' { + ext.args = '--het' + publishDir = [ + path: { "${params.outdir}/variant_calling" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info/variantcalling" }, diff --git a/subworkflows/local/process_vcf.nf b/subworkflows/local/process_vcf.nf new file mode 100644 index 0000000..6f1f45d --- /dev/null +++ b/subworkflows/local/process_vcf.nf @@ -0,0 +1,32 @@ +// +// Call vcftools to process VCF files +// + +include { VCFTOOLS as VCFTOOLS_SITE_PI } from '../../modules/nf-core/vcftools/main' +include { VCFTOOLS as VCFTOOLS_HET } from '../../modules/nf-core/vcftools/main' + +workflow PROCESS_VCF { + take: + vcf // [ val(meta), vcf ] + + main: + ch_versions = Channel.empty() + + // call vcftools for per site nucleotide diversity + VCFTOOLS_SITE_PI( + vcf, [], [] + ) + ch_versions = ch_versions.mix( VCFTOOLS_SITE_PI.out.versions ) + + // call vcftools to calculate for heterozygosity + VCFTOOLS_HET( + vcf, [], [] + ) + ch_versions = ch_versions.mix( VCFTOOLS_HET.out.versions ) + + emit: + versions = ch_versions // channel: [ versions.yml ] + stite_pi = VCFTOOLS_SITE_PI.out.sites_pi // [ meta, site_pi ] + heterozygosity = VCFTOOLS_HET.out.heterozygosity // [ meta, heterozygosity ] + +} diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 82267f7..bfbfa28 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -53,6 +53,7 @@ include { INPUT_CHECK } from '../subworkflows/local/input_check' include { INPUT_MERGE } from '../subworkflows/local/input_merge' include { INPUT_FILTER_SPLIT } from '../subworkflows/local/input_filter_split' include { DEEPVARIANT_CALLER } from '../subworkflows/local/deepvariant_caller' +include { PROCESS_VCF } from '../subworkflows/local/process_vcf' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -114,7 +115,7 @@ workflow VARIANTCALLING { INPUT_CHECK ( ch_input ) - ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) + ch_versions = ch_versions.mix( INPUT_CHECK.out.versions ) // // SUBWORKFLOW: merge the input reads by sample name @@ -124,7 +125,7 @@ workflow VARIANTCALLING { ch_index, INPUT_CHECK.out.reads, ) - ch_versions = ch_versions.mix(INPUT_MERGE.out.versions) + ch_versions = ch_versions.mix( INPUT_MERGE.out.versions ) // @@ -136,7 +137,7 @@ workflow VARIANTCALLING { ch_interval, split_fasta_cutoff ) - ch_versions = ch_versions.mix(INPUT_FILTER_SPLIT.out.versions) + ch_versions = ch_versions.mix( INPUT_FILTER_SPLIT.out.versions ) // // SUBWORKFLOW: call deepvariant @@ -144,7 +145,24 @@ workflow VARIANTCALLING { DEEPVARIANT_CALLER ( INPUT_FILTER_SPLIT.out.reads_fasta ) - ch_versions = ch_versions.mix(DEEPVARIANT_CALLER.out.versions) + ch_versions = ch_versions.mix( DEEPVARIANT_CALLER.out.versions ) + + // + // combine all VCF files together + // + DEEPVARIANT_CALLER.out.gvcf + .map{ meta, gvcf -> [ [ id: gvcf.baseName ], gvcf ] } + .concat( + DEEPVARIANT_CALLER.out.vcf + .map{ meta, vcf -> [ [ id: vcf.baseName ], vcf ] } + ) + .set{ all_vcf } + + // + // process VCF output files + // + PROCESS_VCF( all_vcf ) + ch_versions = ch_versions.mix( PROCESS_VCF.out.versions ) // // MODULE: Combine different version together From 27a04a89c6b4d3b861195cdc45124075da24aae6 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 7 Nov 2023 21:42:47 +0000 Subject: [PATCH 068/123] Patch vcftools modules to allow an input for position files Add include_positions and exclude_positions for the whole workflow. --- conf/modules.config | 5 +++ modules.json | 3 +- modules/nf-core/vcftools/main.nf | 8 ++++ modules/nf-core/vcftools/meta.yml | 5 ++- modules/nf-core/vcftools/vcftools.diff | 51 ++++++++++++++++++++++++++ nextflow.config | 2 + nextflow_schema.json | 8 ++++ subworkflows/local/process_vcf.nf | 7 ++-- workflows/variantcalling.nf | 14 ++++++- 9 files changed, 95 insertions(+), 8 deletions(-) create mode 100644 modules/nf-core/vcftools/vcftools.diff diff --git a/conf/modules.config b/conf/modules.config index 660809c..d9a6ab6 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -53,6 +53,11 @@ process { withName: '.*:PROCESS_VCF:VCFTOOLS_SITE_PI' { ext.args = '--site-pi' + if( params.include_positions ){ + ext.args += ' --positions' + } else if ( params.exclude_positions ){ + ext.args += ' --exclude-positions' + } publishDir = [ path: { "${params.outdir}/variant_calling" }, mode: params.publish_dir_mode, diff --git a/modules.json b/modules.json index 3ce8a13..1ab14cf 100644 --- a/modules.json +++ b/modules.json @@ -49,7 +49,8 @@ "vcftools": { "branch": "master", "git_sha": "8fc1d24c710ebe1d5de0f2447ec9439fd3d9d66a", - "installed_by": ["modules"] + "installed_by": ["modules"], + "patch": "modules/nf-core/vcftools/vcftools.diff" } } } diff --git a/modules/nf-core/vcftools/main.nf b/modules/nf-core/vcftools/main.nf index 0153a60..10fd8d7 100644 --- a/modules/nf-core/vcftools/main.nf +++ b/modules/nf-core/vcftools/main.nf @@ -15,6 +15,7 @@ process VCFTOOLS { tuple val(meta), path(variant_file) path bed path diff_variant_file + path positions_file output: tuple val(meta), path("*.vcf") , optional:true, emit: vcf @@ -103,6 +104,12 @@ process VCFTOOLS { args_list.removeIf { it.contains('--gzdiff') } args_list.removeIf { it.contains('--diff-bcf') } + def positions_arg = (args.contains('--positions')) ? "--positions ${positions_file}" + : (args.contains('--exclude-positions')) ? "--exclude-positions ${positions_file}" + : '' + args_list.removeIf { it.contains('--positions') } + args_list.removeIf { it.contains('--exclude-positions') } + def input_file = ("$variant_file".endsWith(".vcf")) ? "--vcf ${variant_file}" : ("$variant_file".endsWith(".vcf.gz")) ? "--gzvcf ${variant_file}" : ("$variant_file".endsWith(".bcf")) ? "--bcf ${variant_file}" : '' @@ -113,6 +120,7 @@ process VCFTOOLS { --out $prefix \\ ${args_list.join(' ')} \\ $bed_arg \\ + $positions_arg \\ $diff_variant_arg cat <<-END_VERSIONS > versions.yml diff --git a/modules/nf-core/vcftools/meta.yml b/modules/nf-core/vcftools/meta.yml index f361db4..0166bd6 100644 --- a/modules/nf-core/vcftools/meta.yml +++ b/modules/nf-core/vcftools/meta.yml @@ -21,9 +21,10 @@ input: - bed: type: file description: bed file which can be used with different arguments in vcftools (optional) - - diff_variant_file: + - positions_file: type: file - description: secondary variant file which can be used with the 'diff' suite of tools (optional) + description: positions file to include or exclude a set of sites on the basis of a list of positions (optional) + output: - meta: type: map diff --git a/modules/nf-core/vcftools/vcftools.diff b/modules/nf-core/vcftools/vcftools.diff new file mode 100644 index 0000000..2e96489 --- /dev/null +++ b/modules/nf-core/vcftools/vcftools.diff @@ -0,0 +1,51 @@ +Changes in module 'nf-core/vcftools' +--- modules/nf-core/vcftools/meta.yml ++++ modules/nf-core/vcftools/meta.yml +@@ -21,9 +21,10 @@ + - bed: + type: file + description: bed file which can be used with different arguments in vcftools (optional) +- - diff_variant_file: +- type: file +- description: secondary variant file which can be used with the 'diff' suite of tools (optional) ++ - positions_file: ++ type: file ++ description: positions file to include or exclude a set of sites on the basis of a list of positions (optional) ++ + output: + - meta: + type: map + +--- modules/nf-core/vcftools/main.nf ++++ modules/nf-core/vcftools/main.nf +@@ -15,6 +15,7 @@ + tuple val(meta), path(variant_file) + path bed + path diff_variant_file ++ path positions_file + + output: + tuple val(meta), path("*.vcf") , optional:true, emit: vcf +@@ -103,6 +104,12 @@ + args_list.removeIf { it.contains('--gzdiff') } + args_list.removeIf { it.contains('--diff-bcf') } + ++ def positions_arg = (args.contains('--positions')) ? "--positions ${positions_file}" ++ : (args.contains('--exclude-positions')) ? "--exclude-positions ${positions_file}" ++ : '' ++ args_list.removeIf { it.contains('--positions') } ++ args_list.removeIf { it.contains('--exclude-positions') } ++ + def input_file = ("$variant_file".endsWith(".vcf")) ? "--vcf ${variant_file}" : + ("$variant_file".endsWith(".vcf.gz")) ? "--gzvcf ${variant_file}" : + ("$variant_file".endsWith(".bcf")) ? "--bcf ${variant_file}" : '' +@@ -113,6 +120,7 @@ + --out $prefix \\ + ${args_list.join(' ')} \\ + $bed_arg \\ ++ $positions_arg \\ + $diff_variant_arg + + cat <<-END_VERSIONS > versions.yml + +************************************************************ diff --git a/nextflow.config b/nextflow.config index 399b382..90da5d8 100644 --- a/nextflow.config +++ b/nextflow.config @@ -14,6 +14,8 @@ params { fasta = null fai = null interval = null + include_positions = null + exclude_positions = null split_fasta_cutoff = 100000 // Boilerplate options diff --git a/nextflow_schema.json b/nextflow_schema.json index d40c501..e035219 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -60,6 +60,14 @@ "default": 100000, "hidden": true, "description": "The minimum fasta file size when splitting the input fasta file by sequence." + }, + "include_positions": { + "type": "string", + "description": "Path to a file with a set of sites on the basis of a list of positions to include, Each line of the input file should contain a (tab-separated) chromosome and position." + }, + "exclude_positions": { + "type": "string", + "description": "Path to a file with a set of sites on the basis of a list of positions to exclude,Each line of the input file should contain a (tab-separated) chromosome and position." } }, "required": ["fasta"] diff --git a/subworkflows/local/process_vcf.nf b/subworkflows/local/process_vcf.nf index 6f1f45d..8713627 100644 --- a/subworkflows/local/process_vcf.nf +++ b/subworkflows/local/process_vcf.nf @@ -7,20 +7,21 @@ include { VCFTOOLS as VCFTOOLS_HET } from '../../modules/nf-core/vcftool workflow PROCESS_VCF { take: - vcf // [ val(meta), vcf ] + vcf // [ val(meta), vcf ] + site_pi_positions // path to positions file to include or exclude main: ch_versions = Channel.empty() // call vcftools for per site nucleotide diversity VCFTOOLS_SITE_PI( - vcf, [], [] + vcf, [], [], site_pi_positions ) ch_versions = ch_versions.mix( VCFTOOLS_SITE_PI.out.versions ) // call vcftools to calculate for heterozygosity VCFTOOLS_HET( - vcf, [], [] + vcf, [], [], [] ) ch_versions = ch_versions.mix( VCFTOOLS_HET.out.versions ) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index bfbfa28..a54d356 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -10,7 +10,7 @@ def summary_params = NfcoreSchema.paramsSummaryMap(workflow, params) WorkflowVariantcalling.initialise(params, log) // Check input path parameters to see if they exist -def checkPathParamList = [ params.input, params.fasta, params.fai, params.interval ] +def checkPathParamList = [ params.input, params.fasta, params.fai, params.interval, params.include_positions, params.exclude_positions ] for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } // Check mandatory parameters @@ -34,6 +34,16 @@ if (params.interval){ ch_interval = Channel.fromPath(params.interval) } else { c if (params.split_fasta_cutoff ) { split_fasta_cutoff = params.split_fasta_cutoff } else { split_fasta_cutoff = 100000 } +if ( (params.include_positions) && (params.exclude_positions) ){ + exit 1, 'Only one positions file can be given to include or exclude!' +}else if (params.include_positions){ + ch_positions = Channel.fromPath(params.include_positions) +} else if (params.exclude_positions){ + ch_positions = Channel.fromPath(params.exclude_positions) +} else { + ch_positions = Channel.empty() +} + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ CONFIG FILES @@ -161,7 +171,7 @@ workflow VARIANTCALLING { // // process VCF output files // - PROCESS_VCF( all_vcf ) + PROCESS_VCF( all_vcf, ch_positions ) ch_versions = ch_versions.mix( PROCESS_VCF.out.versions ) // From 1feb4d81ecac9e3f77f7711590c172238cbbf951 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 7 Nov 2023 21:51:19 +0000 Subject: [PATCH 069/123] remove trailing white spaces --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index d9a6ab6..cb326e7 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -57,7 +57,7 @@ process { ext.args += ' --positions' } else if ( params.exclude_positions ){ ext.args += ' --exclude-positions' - } + } publishDir = [ path: { "${params.outdir}/variant_calling" }, mode: params.publish_dir_mode, From 3de0d8a679d85b0a06963b22b6fcd4d965147fd8 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 8 Nov 2023 11:13:57 +0000 Subject: [PATCH 070/123] Set the positions to an empty list when no file provided. --- workflows/variantcalling.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index a54d356..4a59fe4 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -41,7 +41,7 @@ if ( (params.include_positions) && (params.exclude_positions) ){ } else if (params.exclude_positions){ ch_positions = Channel.fromPath(params.exclude_positions) } else { - ch_positions = Channel.empty() + ch_positions = [] } /* From a2128daca3e75f780deac461a9c00cc2812c5cd4 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 9 Nov 2023 11:23:50 +0000 Subject: [PATCH 071/123] re-patch vcftools modules --- modules/nf-core/vcftools/main.nf | 4 ++-- modules/nf-core/vcftools/meta.yml | 3 +++ modules/nf-core/vcftools/vcftools.diff | 17 ++++++++--------- 3 files changed, 13 insertions(+), 11 deletions(-) diff --git a/modules/nf-core/vcftools/main.nf b/modules/nf-core/vcftools/main.nf index 10fd8d7..9ef8462 100644 --- a/modules/nf-core/vcftools/main.nf +++ b/modules/nf-core/vcftools/main.nf @@ -120,8 +120,8 @@ process VCFTOOLS { --out $prefix \\ ${args_list.join(' ')} \\ $bed_arg \\ - $positions_arg \\ - $diff_variant_arg + $diff_variant_arg \\ + $positions_arg cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/nf-core/vcftools/meta.yml b/modules/nf-core/vcftools/meta.yml index 0166bd6..fb9551f 100644 --- a/modules/nf-core/vcftools/meta.yml +++ b/modules/nf-core/vcftools/meta.yml @@ -21,6 +21,9 @@ input: - bed: type: file description: bed file which can be used with different arguments in vcftools (optional) + - diff_variant_file: + type: file + description: secondary variant file which can be used with the 'diff' suite of tools (optional) - positions_file: type: file description: positions file to include or exclude a set of sites on the basis of a list of positions (optional) diff --git a/modules/nf-core/vcftools/vcftools.diff b/modules/nf-core/vcftools/vcftools.diff index 2e96489..3343a7d 100644 --- a/modules/nf-core/vcftools/vcftools.diff +++ b/modules/nf-core/vcftools/vcftools.diff @@ -1,13 +1,10 @@ Changes in module 'nf-core/vcftools' --- modules/nf-core/vcftools/meta.yml +++ modules/nf-core/vcftools/meta.yml -@@ -21,9 +21,10 @@ - - bed: +@@ -24,6 +24,10 @@ + - diff_variant_file: type: file - description: bed file which can be used with different arguments in vcftools (optional) -- - diff_variant_file: -- type: file -- description: secondary variant file which can be used with the 'diff' suite of tools (optional) + description: secondary variant file which can be used with the 'diff' suite of tools (optional) + - positions_file: + type: file + description: positions file to include or exclude a set of sites on the basis of a list of positions (optional) @@ -39,13 +36,15 @@ Changes in module 'nf-core/vcftools' def input_file = ("$variant_file".endsWith(".vcf")) ? "--vcf ${variant_file}" : ("$variant_file".endsWith(".vcf.gz")) ? "--gzvcf ${variant_file}" : ("$variant_file".endsWith(".bcf")) ? "--bcf ${variant_file}" : '' -@@ -113,6 +120,7 @@ +@@ -113,7 +120,8 @@ --out $prefix \\ ${args_list.join(' ')} \\ $bed_arg \\ -+ $positions_arg \\ - $diff_variant_arg +- $diff_variant_arg ++ $diff_variant_arg \\ ++ $positions_arg cat <<-END_VERSIONS > versions.yml + "${task.process}": ************************************************************ From c74f90ee08c95fd1f20e60d92dde0b4522949198 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 10 Nov 2023 10:19:03 +0000 Subject: [PATCH 072/123] update vcftools module patch --- modules/nf-core/vcftools/main.nf | 17 ++++---- modules/nf-core/vcftools/meta.yml | 4 -- modules/nf-core/vcftools/vcftools.diff | 54 +++++++------------------- subworkflows/local/process_vcf.nf | 4 +- 4 files changed, 22 insertions(+), 57 deletions(-) diff --git a/modules/nf-core/vcftools/main.nf b/modules/nf-core/vcftools/main.nf index 9ef8462..b36c429 100644 --- a/modules/nf-core/vcftools/main.nf +++ b/modules/nf-core/vcftools/main.nf @@ -15,7 +15,6 @@ process VCFTOOLS { tuple val(meta), path(variant_file) path bed path diff_variant_file - path positions_file output: tuple val(meta), path("*.vcf") , optional:true, emit: vcf @@ -92,10 +91,15 @@ process VCFTOOLS { def bed_arg = (args.contains('--bed')) ? "--bed ${bed}" : (args.contains('--exclude-bed')) ? "--exclude-bed ${bed}" : - (args.contains('--hapcount')) ? "--hapcount ${bed}" : '' + (args.contains('--hapcount')) ? "--hapcount ${bed}" : + (args.contains('--positions')) ? "--positions ${bed}" : + (args.contains('--exclude-positions')) ? "--exclude-positions ${bed}" : '' + args_list.removeIf { it.contains('--bed') } args_list.removeIf { it.contains('--exclude-bed') } args_list.removeIf { it.contains('--hapcount') } + args_list.removeIf { it.contains('--positions') } + args_list.removeIf { it.contains('--exclude-positions') } def diff_variant_arg = (args.contains('--diff')) ? "--diff ${diff_variant_file}" : (args.contains('--gzdiff')) ? "--gzdiff ${diff_variant_file}" : @@ -104,12 +108,6 @@ process VCFTOOLS { args_list.removeIf { it.contains('--gzdiff') } args_list.removeIf { it.contains('--diff-bcf') } - def positions_arg = (args.contains('--positions')) ? "--positions ${positions_file}" - : (args.contains('--exclude-positions')) ? "--exclude-positions ${positions_file}" - : '' - args_list.removeIf { it.contains('--positions') } - args_list.removeIf { it.contains('--exclude-positions') } - def input_file = ("$variant_file".endsWith(".vcf")) ? "--vcf ${variant_file}" : ("$variant_file".endsWith(".vcf.gz")) ? "--gzvcf ${variant_file}" : ("$variant_file".endsWith(".bcf")) ? "--bcf ${variant_file}" : '' @@ -120,8 +118,7 @@ process VCFTOOLS { --out $prefix \\ ${args_list.join(' ')} \\ $bed_arg \\ - $diff_variant_arg \\ - $positions_arg + $diff_variant_arg cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/nf-core/vcftools/meta.yml b/modules/nf-core/vcftools/meta.yml index fb9551f..f361db4 100644 --- a/modules/nf-core/vcftools/meta.yml +++ b/modules/nf-core/vcftools/meta.yml @@ -24,10 +24,6 @@ input: - diff_variant_file: type: file description: secondary variant file which can be used with the 'diff' suite of tools (optional) - - positions_file: - type: file - description: positions file to include or exclude a set of sites on the basis of a list of positions (optional) - output: - meta: type: map diff --git a/modules/nf-core/vcftools/vcftools.diff b/modules/nf-core/vcftools/vcftools.diff index 3343a7d..89e2eae 100644 --- a/modules/nf-core/vcftools/vcftools.diff +++ b/modules/nf-core/vcftools/vcftools.diff @@ -1,50 +1,22 @@ Changes in module 'nf-core/vcftools' ---- modules/nf-core/vcftools/meta.yml -+++ modules/nf-core/vcftools/meta.yml -@@ -24,6 +24,10 @@ - - diff_variant_file: - type: file - description: secondary variant file which can be used with the 'diff' suite of tools (optional) -+ - positions_file: -+ type: file -+ description: positions file to include or exclude a set of sites on the basis of a list of positions (optional) -+ - output: - - meta: - type: map - --- modules/nf-core/vcftools/main.nf +++ modules/nf-core/vcftools/main.nf -@@ -15,6 +15,7 @@ - tuple val(meta), path(variant_file) - path bed - path diff_variant_file -+ path positions_file - - output: - tuple val(meta), path("*.vcf") , optional:true, emit: vcf -@@ -103,6 +104,12 @@ - args_list.removeIf { it.contains('--gzdiff') } - args_list.removeIf { it.contains('--diff-bcf') } +@@ -91,10 +91,15 @@ -+ def positions_arg = (args.contains('--positions')) ? "--positions ${positions_file}" -+ : (args.contains('--exclude-positions')) ? "--exclude-positions ${positions_file}" -+ : '' + def bed_arg = (args.contains('--bed')) ? "--bed ${bed}" : + (args.contains('--exclude-bed')) ? "--exclude-bed ${bed}" : +- (args.contains('--hapcount')) ? "--hapcount ${bed}" : '' ++ (args.contains('--hapcount')) ? "--hapcount ${bed}" : ++ (args.contains('--positions')) ? "--positions ${bed}" : ++ (args.contains('--exclude-positions')) ? "--exclude-positions ${bed}" : '' ++ + args_list.removeIf { it.contains('--bed') } + args_list.removeIf { it.contains('--exclude-bed') } + args_list.removeIf { it.contains('--hapcount') } + args_list.removeIf { it.contains('--positions') } + args_list.removeIf { it.contains('--exclude-positions') } -+ - def input_file = ("$variant_file".endsWith(".vcf")) ? "--vcf ${variant_file}" : - ("$variant_file".endsWith(".vcf.gz")) ? "--gzvcf ${variant_file}" : - ("$variant_file".endsWith(".bcf")) ? "--bcf ${variant_file}" : '' -@@ -113,7 +120,8 @@ - --out $prefix \\ - ${args_list.join(' ')} \\ - $bed_arg \\ -- $diff_variant_arg -+ $diff_variant_arg \\ -+ $positions_arg - cat <<-END_VERSIONS > versions.yml - "${task.process}": + def diff_variant_arg = (args.contains('--diff')) ? "--diff ${diff_variant_file}" : + (args.contains('--gzdiff')) ? "--gzdiff ${diff_variant_file}" : ************************************************************ diff --git a/subworkflows/local/process_vcf.nf b/subworkflows/local/process_vcf.nf index 8713627..432bbf9 100644 --- a/subworkflows/local/process_vcf.nf +++ b/subworkflows/local/process_vcf.nf @@ -15,13 +15,13 @@ workflow PROCESS_VCF { // call vcftools for per site nucleotide diversity VCFTOOLS_SITE_PI( - vcf, [], [], site_pi_positions + vcf, site_pi_positions, [] ) ch_versions = ch_versions.mix( VCFTOOLS_SITE_PI.out.versions ) // call vcftools to calculate for heterozygosity VCFTOOLS_HET( - vcf, [], [], [] + vcf, [], [] ) ch_versions = ch_versions.mix( VCFTOOLS_HET.out.versions ) From e8a24cf3c49410be8b312bc120589f9600656a6b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 13 Nov 2023 16:27:46 +0000 Subject: [PATCH 073/123] not run vcftools on gvcf files --- workflows/variantcalling.nf | 14 +++++--------- 1 file changed, 5 insertions(+), 9 deletions(-) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 4a59fe4..c88480f 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -158,20 +158,16 @@ workflow VARIANTCALLING { ch_versions = ch_versions.mix( DEEPVARIANT_CALLER.out.versions ) // - // combine all VCF files together + // convert VCF channel meta id // - DEEPVARIANT_CALLER.out.gvcf - .map{ meta, gvcf -> [ [ id: gvcf.baseName ], gvcf ] } - .concat( - DEEPVARIANT_CALLER.out.vcf - .map{ meta, vcf -> [ [ id: vcf.baseName ], vcf ] } - ) - .set{ all_vcf } + DEEPVARIANT_CALLER.out.vcf + .map{ meta, vcf -> [ [ id: vcf.baseName ], vcf ] } + .set{ vcf } // // process VCF output files // - PROCESS_VCF( all_vcf, ch_positions ) + PROCESS_VCF( vcf, ch_positions ) ch_versions = ch_versions.mix( PROCESS_VCF.out.versions ) // From 1882fef805d8567142b05a054e2149aca685c0d0 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 16 Nov 2023 09:48:52 +0000 Subject: [PATCH 074/123] Update module vcftools to add one more keywords to minimum 3. --- modules/nf-core/vcftools/meta.yml | 1 + modules/nf-core/vcftools/vcftools.diff | 11 +++++++++++ 2 files changed, 12 insertions(+) diff --git a/modules/nf-core/vcftools/meta.yml b/modules/nf-core/vcftools/meta.yml index f361db4..09ad590 100644 --- a/modules/nf-core/vcftools/meta.yml +++ b/modules/nf-core/vcftools/meta.yml @@ -1,6 +1,7 @@ name: vcftools description: A set of tools written in Perl and C++ for working with VCF files keywords: + - VCFtools - VCF - sort tools: diff --git a/modules/nf-core/vcftools/vcftools.diff b/modules/nf-core/vcftools/vcftools.diff index 89e2eae..fd2e9ec 100644 --- a/modules/nf-core/vcftools/vcftools.diff +++ b/modules/nf-core/vcftools/vcftools.diff @@ -1,4 +1,15 @@ Changes in module 'nf-core/vcftools' +--- modules/nf-core/vcftools/meta.yml ++++ modules/nf-core/vcftools/meta.yml +@@ -1,6 +1,7 @@ + name: vcftools + description: A set of tools written in Perl and C++ for working with VCF files + keywords: ++ - VCFtools + - VCF + - sort + tools: + --- modules/nf-core/vcftools/main.nf +++ modules/nf-core/vcftools/main.nf @@ -91,10 +91,15 @@ From 45de788c5d25833c974f1acc392d464d92ce15cc Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 17 Nov 2023 12:37:35 +0000 Subject: [PATCH 075/123] copy align_pacbio subworkflow from readmapping pipleline dev branch, 093b313e29ed68480d81d796cc1609536518ee5a and install all the related modules. --- modules.json | 40 ++++++++++ modules/local/pacbio_filter.nf | 30 +++++++ modules/nf-core/blast/blastn/environment.yml | 7 ++ modules/nf-core/blast/blastn/main.nf | 57 ++++++++++++++ modules/nf-core/blast/blastn/meta.yml | 55 +++++++++++++ .../nf-core/blast/blastn/tests/main.nf.test | 71 +++++++++++++++++ .../blast/blastn/tests/nextflow.config | 5 ++ modules/nf-core/blast/blastn/tests/tags.yml | 2 + modules/nf-core/gunzip/environment.yml | 7 ++ modules/nf-core/gunzip/main.nf | 48 ++++++++++++ modules/nf-core/gunzip/meta.yml | 39 ++++++++++ modules/nf-core/gunzip/tests/main.nf.test | 35 +++++++++ .../nf-core/gunzip/tests/main.nf.test.snap | 31 ++++++++ modules/nf-core/gunzip/tests/tags.yml | 2 + .../nf-core/minimap2/align/environment.yml | 8 ++ modules/nf-core/minimap2/align/main.nf | 48 ++++++++++++ modules/nf-core/minimap2/align/meta.yml | 75 ++++++++++++++++++ .../nf-core/samtools/collate/environment.yml | 7 ++ modules/nf-core/samtools/collate/main.nf | 46 +++++++++++ modules/nf-core/samtools/collate/meta.yml | 43 ++++++++++ .../nf-core/samtools/fasta/environment.yml | 7 ++ modules/nf-core/samtools/fasta/main.nf | 44 +++++++++++ modules/nf-core/samtools/fasta/meta.yml | 60 ++++++++++++++ .../nf-core/samtools/flagstat/environment.yml | 7 ++ modules/nf-core/samtools/flagstat/main.nf | 46 +++++++++++ modules/nf-core/samtools/flagstat/meta.yml | 51 ++++++++++++ .../samtools/flagstat/tests/main.nf.test | 35 +++++++++ .../samtools/flagstat/tests/main.nf.test.snap | 16 ++++ .../nf-core/samtools/flagstat/tests/tags.yml | 2 + .../nf-core/samtools/idxstats/environment.yml | 7 ++ modules/nf-core/samtools/idxstats/main.nf | 48 ++++++++++++ modules/nf-core/samtools/idxstats/meta.yml | 52 +++++++++++++ .../samtools/idxstats/tests/main.nf.test | 35 +++++++++ .../samtools/idxstats/tests/main.nf.test.snap | 16 ++++ .../nf-core/samtools/idxstats/tests/tags.yml | 2 + .../nf-core/samtools/stats/environment.yml | 7 ++ modules/nf-core/samtools/stats/main.nf | 49 ++++++++++++ modules/nf-core/samtools/stats/meta.yml | 63 +++++++++++++++ .../nf-core/samtools/stats/tests/main.nf.test | 78 +++++++++++++++++++ .../samtools/stats/tests/main.nf.test.snap | 64 +++++++++++++++ modules/nf-core/samtools/stats/tests/tags.yml | 2 + subworkflows/local/align_pacbio.nf | 70 +++++++++++++++++ subworkflows/local/convert_stats.nf | 54 +++++++++++++ subworkflows/local/filter_pacbio.nf | 77 ++++++++++++++++++ 44 files changed, 1548 insertions(+) create mode 100644 modules/local/pacbio_filter.nf create mode 100644 modules/nf-core/blast/blastn/environment.yml create mode 100644 modules/nf-core/blast/blastn/main.nf create mode 100644 modules/nf-core/blast/blastn/meta.yml create mode 100644 modules/nf-core/blast/blastn/tests/main.nf.test create mode 100644 modules/nf-core/blast/blastn/tests/nextflow.config create mode 100644 modules/nf-core/blast/blastn/tests/tags.yml create mode 100644 modules/nf-core/gunzip/environment.yml create mode 100644 modules/nf-core/gunzip/main.nf create mode 100644 modules/nf-core/gunzip/meta.yml create mode 100644 modules/nf-core/gunzip/tests/main.nf.test create mode 100644 modules/nf-core/gunzip/tests/main.nf.test.snap create mode 100644 modules/nf-core/gunzip/tests/tags.yml create mode 100644 modules/nf-core/minimap2/align/environment.yml create mode 100644 modules/nf-core/minimap2/align/main.nf create mode 100644 modules/nf-core/minimap2/align/meta.yml create mode 100644 modules/nf-core/samtools/collate/environment.yml create mode 100644 modules/nf-core/samtools/collate/main.nf create mode 100644 modules/nf-core/samtools/collate/meta.yml create mode 100644 modules/nf-core/samtools/fasta/environment.yml create mode 100644 modules/nf-core/samtools/fasta/main.nf create mode 100644 modules/nf-core/samtools/fasta/meta.yml create mode 100644 modules/nf-core/samtools/flagstat/environment.yml create mode 100644 modules/nf-core/samtools/flagstat/main.nf create mode 100644 modules/nf-core/samtools/flagstat/meta.yml create mode 100644 modules/nf-core/samtools/flagstat/tests/main.nf.test create mode 100644 modules/nf-core/samtools/flagstat/tests/main.nf.test.snap create mode 100644 modules/nf-core/samtools/flagstat/tests/tags.yml create mode 100644 modules/nf-core/samtools/idxstats/environment.yml create mode 100644 modules/nf-core/samtools/idxstats/main.nf create mode 100644 modules/nf-core/samtools/idxstats/meta.yml create mode 100644 modules/nf-core/samtools/idxstats/tests/main.nf.test create mode 100644 modules/nf-core/samtools/idxstats/tests/main.nf.test.snap create mode 100644 modules/nf-core/samtools/idxstats/tests/tags.yml create mode 100644 modules/nf-core/samtools/stats/environment.yml create mode 100644 modules/nf-core/samtools/stats/main.nf create mode 100644 modules/nf-core/samtools/stats/meta.yml create mode 100644 modules/nf-core/samtools/stats/tests/main.nf.test create mode 100644 modules/nf-core/samtools/stats/tests/main.nf.test.snap create mode 100644 modules/nf-core/samtools/stats/tests/tags.yml create mode 100644 subworkflows/local/align_pacbio.nf create mode 100644 subworkflows/local/convert_stats.nf create mode 100644 subworkflows/local/filter_pacbio.nf diff --git a/modules.json b/modules.json index 1ab14cf..1e3636d 100644 --- a/modules.json +++ b/modules.json @@ -10,6 +10,11 @@ "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] }, + "blast/blastn": { + "branch": "master", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "installed_by": ["modules"] + }, "cat/cat": { "branch": "master", "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", @@ -25,11 +30,41 @@ "git_sha": "516189e968feb4ebdd9921806988b4c12b4ac2dc", "installed_by": ["modules"] }, + "gunzip": { + "branch": "master", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "installed_by": ["modules"] + }, + "minimap2/align": { + "branch": "master", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "installed_by": ["modules"] + }, + "samtools/collate": { + "branch": "master", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "installed_by": ["modules"] + }, "samtools/faidx": { "branch": "master", "git_sha": "fd742419940e01ba1c5ecb172c3e32ec840662fe", "installed_by": ["modules"] }, + "samtools/fasta": { + "branch": "master", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "installed_by": ["modules"] + }, + "samtools/flagstat": { + "branch": "master", + "git_sha": "63e817de8c617131447192ab2c4e70b4ed4071f7", + "installed_by": ["modules"] + }, + "samtools/idxstats": { + "branch": "master", + "git_sha": "63e817de8c617131447192ab2c4e70b4ed4071f7", + "installed_by": ["modules"] + }, "samtools/merge": { "branch": "master", "git_sha": "e7ce60acc8a33fa17429e966364657a63016e870", @@ -41,6 +76,11 @@ "git_sha": "a0f7be95788366c1923171e358da7d049eb440f9", "installed_by": ["modules"] }, + "samtools/stats": { + "branch": "master", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "installed_by": ["modules"] + }, "samtools/view": { "branch": "master", "git_sha": "3ffae3598260a99e8db3207dead9f73f87f90d1f", diff --git a/modules/local/pacbio_filter.nf b/modules/local/pacbio_filter.nf new file mode 100644 index 0000000..18dd11c --- /dev/null +++ b/modules/local/pacbio_filter.nf @@ -0,0 +1,30 @@ +process PACBIO_FILTER { + tag "$meta.id" + label 'process_single' + + conda "conda-forge::gawk=5.1.0" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/gawk:5.1.0' : + 'quay.io/biocontainers/gawk:5.1.0' }" + + input: + tuple val(meta), path(txt) + + output: + path("*.blocklist"), emit: list + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + pacbio_filter.sh $txt ${prefix}.blocklist + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + GNU Awk: \$(echo \$(awk --version 2>&1) | grep -i awk | sed 's/GNU Awk //; s/,.*//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/blast/blastn/environment.yml b/modules/nf-core/blast/blastn/environment.yml new file mode 100644 index 0000000..cb9b15d --- /dev/null +++ b/modules/nf-core/blast/blastn/environment.yml @@ -0,0 +1,7 @@ +name: blast_blastn +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::blast=2.14.1 diff --git a/modules/nf-core/blast/blastn/main.nf b/modules/nf-core/blast/blastn/main.nf new file mode 100644 index 0000000..e8b96ad --- /dev/null +++ b/modules/nf-core/blast/blastn/main.nf @@ -0,0 +1,57 @@ +process BLAST_BLASTN { + tag "$meta.id" + label 'process_medium' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/blast:2.14.1--pl5321h6f7f691_0': + 'biocontainers/blast:2.14.1--pl5321h6f7f691_0' }" + + input: + tuple val(meta) , path(fasta) + tuple val(meta2), path(db) + + output: + tuple val(meta), path('*.txt'), emit: txt + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def is_compressed = fasta.getExtension() == "gz" ? true : false + def fasta_name = is_compressed ? fasta.getBaseName() : fasta + + """ + if [ "${is_compressed}" == "true" ]; then + gzip -c -d ${fasta} > ${fasta_name} + fi + + DB=`find -L ./ -name "*.nin" | sed 's/\\.nin\$//'` + blastn \\ + -num_threads ${task.cpus} \\ + -db \$DB \\ + -query ${fasta_name} \\ + ${args} \\ + -out ${prefix}.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + blast: \$(blastn -version 2>&1 | sed 's/^.*blastn: //; s/ .*\$//') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch ${prefix}.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + blast: \$(blastn -version 2>&1 | sed 's/^.*blastn: //; s/ .*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/blast/blastn/meta.yml b/modules/nf-core/blast/blastn/meta.yml new file mode 100644 index 0000000..a0d64dd --- /dev/null +++ b/modules/nf-core/blast/blastn/meta.yml @@ -0,0 +1,55 @@ +name: blast_blastn +description: Queries a BLAST DNA database +keywords: + - fasta + - blast + - blastn + - DNA sequence +tools: + - blast: + description: | + BLAST finds regions of similarity between biological sequences. + homepage: https://blast.ncbi.nlm.nih.gov/Blast.cgi + documentation: https://blast.ncbi.nlm.nih.gov/Blast.cgi?CMD=Web&PAGE_TYPE=Blastdocs + doi: 10.1016/S0022-2836(05)80360-2 + licence: ["US-Government-Work"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Input fasta file containing queries sequences + pattern: "*.{fa,fasta,fa.gz,fasta.gz}" + - meta2: + type: map + description: | + Groovy Map containing db information + e.g. [ id:'test2', single_end:false ] + - db: + type: directory + description: Directory containing the blast database + pattern: "*" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - txt: + type: file + description: File containing blastn hits + pattern: "*.txt" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" +maintainers: + - "@joseespinosa" + - "@drpatelh" + - "@vagkaratzas" diff --git a/modules/nf-core/blast/blastn/tests/main.nf.test b/modules/nf-core/blast/blastn/tests/main.nf.test new file mode 100644 index 0000000..1058c81 --- /dev/null +++ b/modules/nf-core/blast/blastn/tests/main.nf.test @@ -0,0 +1,71 @@ +nextflow_process { + + name "Test Process BLAST_BLASTN" + script "../main.nf" + process "BLAST_BLASTN" + config "./nextflow.config" + tag "modules" + tag "modules_nfcore" + tag "blast" + tag "blast/blastn" + + setup { + run("BLAST_MAKEBLASTDB") { + script "../../makeblastdb/main.nf" + process { + """ + input[0] = [ [id:'test2'], file(params.test_data['sarscov2']['genome']['genome_fasta'], checkIfExists: true) ] + """ + } + } + } + + test("Should search for nucleotide hits against a blast db") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ [id:'test'], file(params.test_data['sarscov2']['genome']['genome_fasta'], checkIfExists: true) ] + input[1] = BLAST_MAKEBLASTDB.out.db + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert path(process.out.txt.get(0).get(1)).getText().contains("Query= MT192765.1 Severe acute respiratory syndrome coronavirus 2 isolate") }, + { assert process.out.versions } + ) + } + + } + + test("Should search for zipped nucleotide hits against a blast db") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ [id:'test'], file(params.test_data['sarscov2']['genome']['genome_fasta_gz'], checkIfExists: true) ] + input[1] = BLAST_MAKEBLASTDB.out.db + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert path(process.out.txt.get(0).get(1)).getText().contains("Query= MT192765.1 Severe acute respiratory syndrome coronavirus 2 isolate") }, + { assert process.out.versions } + ) + } + + } + +} diff --git a/modules/nf-core/blast/blastn/tests/nextflow.config b/modules/nf-core/blast/blastn/tests/nextflow.config new file mode 100644 index 0000000..0899289 --- /dev/null +++ b/modules/nf-core/blast/blastn/tests/nextflow.config @@ -0,0 +1,5 @@ +process { + withName: BLAST_MAKEBLASTDB { + ext.args = '-dbtype nucl' + } +} diff --git a/modules/nf-core/blast/blastn/tests/tags.yml b/modules/nf-core/blast/blastn/tests/tags.yml new file mode 100644 index 0000000..b4588ab --- /dev/null +++ b/modules/nf-core/blast/blastn/tests/tags.yml @@ -0,0 +1,2 @@ +blast/blastn: + - modules/nf-core/blast/blastn/** diff --git a/modules/nf-core/gunzip/environment.yml b/modules/nf-core/gunzip/environment.yml new file mode 100644 index 0000000..25910b3 --- /dev/null +++ b/modules/nf-core/gunzip/environment.yml @@ -0,0 +1,7 @@ +name: gunzip +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - conda-forge::sed=4.7 diff --git a/modules/nf-core/gunzip/main.nf b/modules/nf-core/gunzip/main.nf new file mode 100644 index 0000000..468a6f2 --- /dev/null +++ b/modules/nf-core/gunzip/main.nf @@ -0,0 +1,48 @@ +process GUNZIP { + tag "$archive" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'nf-core/ubuntu:20.04' }" + + input: + tuple val(meta), path(archive) + + output: + tuple val(meta), path("$gunzip"), emit: gunzip + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + gunzip = archive.toString() - '.gz' + """ + # Not calling gunzip itself because it creates files + # with the original group ownership rather than the + # default one for that user / the work directory + gzip \\ + -cd \\ + $args \\ + $archive \\ + > $gunzip + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + gunzip: \$(echo \$(gunzip --version 2>&1) | sed 's/^.*(gzip) //; s/ Copyright.*\$//') + END_VERSIONS + """ + + stub: + gunzip = archive.toString() - '.gz' + """ + touch $gunzip + cat <<-END_VERSIONS > versions.yml + "${task.process}": + gunzip: \$(echo \$(gunzip --version 2>&1) | sed 's/^.*(gzip) //; s/ Copyright.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/gunzip/meta.yml b/modules/nf-core/gunzip/meta.yml new file mode 100644 index 0000000..231034f --- /dev/null +++ b/modules/nf-core/gunzip/meta.yml @@ -0,0 +1,39 @@ +name: gunzip +description: Compresses and decompresses files. +keywords: + - gunzip + - compression + - decompression +tools: + - gunzip: + description: | + gzip is a file format and a software application used for file compression and decompression. + documentation: https://www.gnu.org/software/gzip/manual/gzip.html + licence: ["GPL-3.0-or-later"] +input: + - meta: + type: map + description: | + Optional groovy Map containing meta information + e.g. [ id:'test', single_end:false ] + - archive: + type: file + description: File to be compressed/uncompressed + pattern: "*.*" +output: + - gunzip: + type: file + description: Compressed/uncompressed file + pattern: "*.*" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" + - "@jfy133" +maintainers: + - "@joseespinosa" + - "@drpatelh" + - "@jfy133" diff --git a/modules/nf-core/gunzip/tests/main.nf.test b/modules/nf-core/gunzip/tests/main.nf.test new file mode 100644 index 0000000..d031792 --- /dev/null +++ b/modules/nf-core/gunzip/tests/main.nf.test @@ -0,0 +1,35 @@ +nextflow_process { + + name "Test Process GUNZIP" + script "../main.nf" + process "GUNZIP" + tag "gunzip" + tag "modules_nfcore" + tag "modules" + + test("Should run without failures") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ + [], + file(params.test_data['sarscov2']['illumina']['test_1_fastq_gz'], checkIfExists: true) + ] + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot(process.out).match() } + ) + } + + } + +} diff --git a/modules/nf-core/gunzip/tests/main.nf.test.snap b/modules/nf-core/gunzip/tests/main.nf.test.snap new file mode 100644 index 0000000..720fd9f --- /dev/null +++ b/modules/nf-core/gunzip/tests/main.nf.test.snap @@ -0,0 +1,31 @@ +{ + "Should run without failures": { + "content": [ + { + "0": [ + [ + [ + + ], + "test_1.fastq:md5,4161df271f9bfcd25d5845a1e220dbec" + ] + ], + "1": [ + "versions.yml:md5,54376d32aca20e937a4ec26dac228e84" + ], + "gunzip": [ + [ + [ + + ], + "test_1.fastq:md5,4161df271f9bfcd25d5845a1e220dbec" + ] + ], + "versions": [ + "versions.yml:md5,54376d32aca20e937a4ec26dac228e84" + ] + } + ], + "timestamp": "2023-10-17T15:35:37.690477896" + } +} \ No newline at end of file diff --git a/modules/nf-core/gunzip/tests/tags.yml b/modules/nf-core/gunzip/tests/tags.yml new file mode 100644 index 0000000..fd3f691 --- /dev/null +++ b/modules/nf-core/gunzip/tests/tags.yml @@ -0,0 +1,2 @@ +gunzip: + - modules/nf-core/gunzip/** diff --git a/modules/nf-core/minimap2/align/environment.yml b/modules/nf-core/minimap2/align/environment.yml new file mode 100644 index 0000000..60b9a8b --- /dev/null +++ b/modules/nf-core/minimap2/align/environment.yml @@ -0,0 +1,8 @@ +name: minimap2_align +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::minimap2=2.24 + - bioconda::samtools=1.14 diff --git a/modules/nf-core/minimap2/align/main.nf b/modules/nf-core/minimap2/align/main.nf new file mode 100644 index 0000000..fa3ae50 --- /dev/null +++ b/modules/nf-core/minimap2/align/main.nf @@ -0,0 +1,48 @@ +process MINIMAP2_ALIGN { + tag "$meta.id" + label 'process_medium' + + // Note: the versions here need to match the versions used in the mulled container below and minimap2/index + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/mulled-v2-66534bcbb7031a148b13e2ad42583020b9cd25c4:1679e915ddb9d6b4abda91880c4b48857d471bd8-0' : + 'biocontainers/mulled-v2-66534bcbb7031a148b13e2ad42583020b9cd25c4:1679e915ddb9d6b4abda91880c4b48857d471bd8-0' }" + + input: + tuple val(meta), path(reads) + tuple val(meta2), path(reference) + val bam_format + val cigar_paf_format + val cigar_bam + + output: + tuple val(meta), path("*.paf"), optional: true, emit: paf + tuple val(meta), path("*.bam"), optional: true, emit: bam + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def bam_output = bam_format ? "-a | samtools sort | samtools view -@ ${task.cpus} -b -h -o ${prefix}.bam" : "-o ${prefix}.paf" + def cigar_paf = cigar_paf_format && !bam_format ? "-c" : '' + def set_cigar_bam = cigar_bam && bam_format ? "-L" : '' + """ + minimap2 \\ + $args \\ + -t $task.cpus \\ + "${reference ?: reads}" \\ + "$reads" \\ + $cigar_paf \\ + $set_cigar_bam \\ + $bam_output + + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + minimap2: \$(minimap2 --version 2>&1) + END_VERSIONS + """ +} diff --git a/modules/nf-core/minimap2/align/meta.yml b/modules/nf-core/minimap2/align/meta.yml new file mode 100644 index 0000000..408522d --- /dev/null +++ b/modules/nf-core/minimap2/align/meta.yml @@ -0,0 +1,75 @@ +name: minimap2_align +description: A versatile pairwise aligner for genomic and spliced nucleotide sequences +keywords: + - align + - fasta + - fastq + - genome + - paf + - reference +tools: + - minimap2: + description: | + A versatile pairwise aligner for genomic and spliced nucleotide sequences. + homepage: https://github.com/lh3/minimap2 + documentation: https://github.com/lh3/minimap2#uguide + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FASTA or FASTQ files of size 1 and 2 for single-end + and paired-end data, respectively. + - meta2: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'test_ref'] + - reference: + type: file + description: | + Reference database in FASTA format. + - bam_format: + type: boolean + description: Specify that output should be in BAM format + - cigar_paf_format: + type: boolean + description: Specify that output CIGAR should be in PAF format + - cigar_bam: + type: boolean + description: | + Write CIGAR with >65535 ops at the CG tag. This is recommended when + doing XYZ (https://github.com/lh3/minimap2#working-with-65535-cigar-operations) +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - paf: + type: file + description: Alignment in PAF format + pattern: "*.paf" + - bam: + type: file + description: Alignment in BAM format + pattern: "*.bam" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@heuermh" + - "@sofstam" + - "@sateeshperi" + - "@jfy133" +maintainers: + - "@heuermh" + - "@sofstam" + - "@sateeshperi" + - "@jfy133" diff --git a/modules/nf-core/samtools/collate/environment.yml b/modules/nf-core/samtools/collate/environment.yml new file mode 100644 index 0000000..0fb861b --- /dev/null +++ b/modules/nf-core/samtools/collate/environment.yml @@ -0,0 +1,7 @@ +name: samtools_collate +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/collate/main.nf b/modules/nf-core/samtools/collate/main.nf new file mode 100644 index 0000000..38a4daf --- /dev/null +++ b/modules/nf-core/samtools/collate/main.nf @@ -0,0 +1,46 @@ +process SAMTOOLS_COLLATE { + tag "$meta.id" + label 'process_medium' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0': + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(input) + path fasta + + output: + tuple val(meta), path("*.bam"), emit: bam, optional: true + tuple val(meta), path("*.cram"), emit: cram, optional: true + tuple val(meta), path("*.sam"), emit: sam, optional: true + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def reference = fasta ? "--reference ${fasta}" : "" + def extension = args.contains("--output-fmt sam") ? "sam" : + args.contains("--output-fmt bam") ? "bam" : + args.contains("--output-fmt cram") ? "cram" : + "bam" + if ("$input" == "${prefix}.${extension}") error "Input and output names are the same, use \"task.ext.prefix\" to disambiguate!" + """ + samtools \\ + collate \\ + $args \\ + ${reference} \\ + -@ $task.cpus \\ + -o ${prefix}.${extension} \\ + $input + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/collate/meta.yml b/modules/nf-core/samtools/collate/meta.yml new file mode 100644 index 0000000..3c79927 --- /dev/null +++ b/modules/nf-core/samtools/collate/meta.yml @@ -0,0 +1,43 @@ +name: "samtools_collate" +description: shuffles and groups reads together by their names +keywords: + - collate + - bam +tools: + - "samtools": + description: "Tools for dealing with SAM, BAM and CRAM files" + homepage: "http://www.htslib.org" + documentation: "https://www.htslib.org/doc/samtools-collate.html" + tool_dev_url: "https://github.com/samtools/samtools" + doi: "10.1093/bioinformatics/btp352" + licence: "['MIT']" +input: + # Only when we have meta + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - input: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" +output: + #Only when we have meta + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - output: + type: file + description: Collated BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" +authors: + - "@priyanka-surana" +maintainers: + - "@priyanka-surana" diff --git a/modules/nf-core/samtools/fasta/environment.yml b/modules/nf-core/samtools/fasta/environment.yml new file mode 100644 index 0000000..8a82f9e --- /dev/null +++ b/modules/nf-core/samtools/fasta/environment.yml @@ -0,0 +1,7 @@ +name: samtools_fasta +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/fasta/main.nf b/modules/nf-core/samtools/fasta/main.nf new file mode 100644 index 0000000..925ed62 --- /dev/null +++ b/modules/nf-core/samtools/fasta/main.nf @@ -0,0 +1,44 @@ +process SAMTOOLS_FASTA { + tag "$meta.id" + label 'process_low' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(input) + val(interleave) + + output: + tuple val(meta), path("*_{1,2}.fasta.gz") , optional:true, emit: fasta + tuple val(meta), path("*_interleaved.fasta.gz"), optional:true, emit: interleaved + tuple val(meta), path("*_singleton.fasta.gz") , optional:true, emit: singleton + tuple val(meta), path("*_other.fasta.gz") , optional:true, emit: other + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def output = ( interleave && ! meta.single_end ) ? "> ${prefix}_interleaved.fasta.gz" : + meta.single_end ? "-1 ${prefix}_1.fasta.gz -s ${prefix}_singleton.fasta.gz" : + "-1 ${prefix}_1.fasta.gz -2 ${prefix}_2.fasta.gz -s ${prefix}_singleton.fasta.gz" + """ + samtools \\ + fasta \\ + $args \\ + --threads ${task.cpus-1} \\ + -0 ${prefix}_other.fasta.gz \\ + $input \\ + $output + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//' )) + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/fasta/meta.yml b/modules/nf-core/samtools/fasta/meta.yml new file mode 100644 index 0000000..1d07ea1 --- /dev/null +++ b/modules/nf-core/samtools/fasta/meta.yml @@ -0,0 +1,60 @@ +name: "samtools_fasta" +description: Converts a SAM/BAM/CRAM file to FASTA +keywords: + - bam + - sam + - cram + - fasta +tools: + - "samtools": + description: "Tools for dealing with SAM, BAM and CRAM files" + homepage: "http://www.htslib.org" + documentation: "https://www.htslib.org/doc/samtools-fasta.html" + tool_dev_url: "https://github.com/samtools/samtools" + doi: "10.1093/bioinformatics/btp352" + licence: "['MIT']" +input: + # Only when we have meta + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - input: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - interleave: + type: boolean + description: Set true for interleaved fasta files +output: + #Only when we have meta + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - fasta: + type: file + description: Compressed FASTA file(s) with reads with either the READ1 or READ2 flag set in separate files. + pattern: "*_{1,2}.fasta.gz" + - interleaved: + type: file + description: Compressed FASTA file with reads with either the READ1 or READ2 flag set in a combined file. Needs collated input file. + pattern: "*_interleaved.fasta.gz" + - singleton: + type: file + description: Compressed FASTA file with singleton reads + pattern: "*_singleton.fasta.gz" + - other: + type: file + description: Compressed FASTA file with reads with either both READ1 and READ2 flags set or unset + pattern: "*_other.fasta.gz" +authors: + - "@priyanka-surana" +maintainers: + - "@priyanka-surana" diff --git a/modules/nf-core/samtools/flagstat/environment.yml b/modules/nf-core/samtools/flagstat/environment.yml new file mode 100644 index 0000000..22bdb5c --- /dev/null +++ b/modules/nf-core/samtools/flagstat/environment.yml @@ -0,0 +1,7 @@ +name: samtools_flagstat +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/flagstat/main.nf b/modules/nf-core/samtools/flagstat/main.nf new file mode 100644 index 0000000..9dee35a --- /dev/null +++ b/modules/nf-core/samtools/flagstat/main.nf @@ -0,0 +1,46 @@ +process SAMTOOLS_FLAGSTAT { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(bam), path(bai) + + output: + tuple val(meta), path("*.flagstat"), emit: flagstat + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + """ + samtools \\ + flagstat \\ + --threads ${task.cpus} \\ + $bam \\ + > ${prefix}.flagstat + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch ${prefix}.flagstat + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/flagstat/meta.yml b/modules/nf-core/samtools/flagstat/meta.yml new file mode 100644 index 0000000..9799135 --- /dev/null +++ b/modules/nf-core/samtools/flagstat/meta.yml @@ -0,0 +1,51 @@ +name: samtools_flagstat +description: Counts the number of alignments in a BAM/CRAM/SAM file for each FLAG type +keywords: + - stats + - mapping + - counts + - bam + - sam + - cram +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - bai: + type: file + description: Index for BAM/CRAM/SAM file + pattern: "*.{bai,crai,sai}" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - flagstat: + type: file + description: File containing samtools flagstat output + pattern: "*.{flagstat}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" +maintainers: + - "@drpatelh" diff --git a/modules/nf-core/samtools/flagstat/tests/main.nf.test b/modules/nf-core/samtools/flagstat/tests/main.nf.test new file mode 100644 index 0000000..c618de7 --- /dev/null +++ b/modules/nf-core/samtools/flagstat/tests/main.nf.test @@ -0,0 +1,35 @@ +nextflow_process { + + name "Test Process SAMTOOLS_FLAGSTAT" + script "../main.nf" + process "SAMTOOLS_FLAGSTAT" + tag "modules" + tag "modules_nfcore" + tag "samtools/flagstat" + + test("BAM") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ + [ id:'test', single_end:false ], // meta map + file(params.test_data['sarscov2']['illumina']['test_paired_end_sorted_bam'], checkIfExists: true), + file(params.test_data['sarscov2']['illumina']['test_paired_end_sorted_bam_bai'], checkIfExists: true) + ] + """ + } + } + + then { + assertAll ( + { assert process.success }, + { assert snapshot(process.out.flagstat).match() }, + { assert path(process.out.versions.get(0)).getText().contains("samtools") } + ) + } + } +} diff --git a/modules/nf-core/samtools/flagstat/tests/main.nf.test.snap b/modules/nf-core/samtools/flagstat/tests/main.nf.test.snap new file mode 100644 index 0000000..880019f --- /dev/null +++ b/modules/nf-core/samtools/flagstat/tests/main.nf.test.snap @@ -0,0 +1,16 @@ +{ + "BAM": { + "content": [ + [ + [ + { + "id": "test", + "single_end": false + }, + "test.flagstat:md5,4f7ffd1e6a5e85524d443209ac97d783" + ] + ] + ], + "timestamp": "2023-11-14T15:49:22.577133" + } +} \ No newline at end of file diff --git a/modules/nf-core/samtools/flagstat/tests/tags.yml b/modules/nf-core/samtools/flagstat/tests/tags.yml new file mode 100644 index 0000000..2d2b725 --- /dev/null +++ b/modules/nf-core/samtools/flagstat/tests/tags.yml @@ -0,0 +1,2 @@ +samtools/flagstat: + - modules/nf-core/samtools/flagstat/** diff --git a/modules/nf-core/samtools/idxstats/environment.yml b/modules/nf-core/samtools/idxstats/environment.yml new file mode 100644 index 0000000..89bd272 --- /dev/null +++ b/modules/nf-core/samtools/idxstats/environment.yml @@ -0,0 +1,7 @@ +name: samtools_idxstats +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/idxstats/main.nf b/modules/nf-core/samtools/idxstats/main.nf new file mode 100644 index 0000000..b22d084 --- /dev/null +++ b/modules/nf-core/samtools/idxstats/main.nf @@ -0,0 +1,48 @@ +process SAMTOOLS_IDXSTATS { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(bam), path(bai) + + output: + tuple val(meta), path("*.idxstats"), emit: idxstats + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + + """ + samtools \\ + idxstats \\ + --threads ${task.cpus-1} \\ + $bam \\ + > ${prefix}.idxstats + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + + """ + touch ${prefix}.idxstats + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/idxstats/meta.yml b/modules/nf-core/samtools/idxstats/meta.yml new file mode 100644 index 0000000..344e92a --- /dev/null +++ b/modules/nf-core/samtools/idxstats/meta.yml @@ -0,0 +1,52 @@ +name: samtools_idxstats +description: Reports alignment summary statistics for a BAM/CRAM/SAM file +keywords: + - stats + - mapping + - counts + - chromosome + - bam + - sam + - cram +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - bam: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - bai: + type: file + description: Index for BAM/CRAM/SAM file + pattern: "*.{bai,crai,sai}" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - idxstats: + type: file + description: File containing samtools idxstats output + pattern: "*.{idxstats}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" +maintainers: + - "@drpatelh" diff --git a/modules/nf-core/samtools/idxstats/tests/main.nf.test b/modules/nf-core/samtools/idxstats/tests/main.nf.test new file mode 100644 index 0000000..0174a9e --- /dev/null +++ b/modules/nf-core/samtools/idxstats/tests/main.nf.test @@ -0,0 +1,35 @@ +nextflow_process { + + name "Test Process SAMTOOLS_IDXSTATS" + script "../main.nf" + process "SAMTOOLS_IDXSTATS" + tag "modules" + tag "modules_nfcore" + tag "samtools/idxstats" + + test("BAM") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ + [ id:'test', single_end:false ], // meta map + file(params.test_data['sarscov2']['illumina']['test_paired_end_sorted_bam'], checkIfExists: true), + file(params.test_data['sarscov2']['illumina']['test_paired_end_sorted_bam_bai'], checkIfExists: true) + ] + """ + } + } + + then { + assertAll ( + { assert process.success }, + { assert snapshot(process.out.idxstats).match() }, + { assert path(process.out.versions.get(0)).getText().contains("samtools") } + ) + } + } +} diff --git a/modules/nf-core/samtools/idxstats/tests/main.nf.test.snap b/modules/nf-core/samtools/idxstats/tests/main.nf.test.snap new file mode 100644 index 0000000..4c6c12b --- /dev/null +++ b/modules/nf-core/samtools/idxstats/tests/main.nf.test.snap @@ -0,0 +1,16 @@ +{ + "BAM": { + "content": [ + [ + [ + { + "id": "test", + "single_end": false + }, + "test.idxstats:md5,df60a8c8d6621100d05178c93fb053a2" + ] + ] + ], + "timestamp": "2023-11-14T15:52:19.875194" + } +} \ No newline at end of file diff --git a/modules/nf-core/samtools/idxstats/tests/tags.yml b/modules/nf-core/samtools/idxstats/tests/tags.yml new file mode 100644 index 0000000..d3057c6 --- /dev/null +++ b/modules/nf-core/samtools/idxstats/tests/tags.yml @@ -0,0 +1,2 @@ +samtools/idxstats: + - modules/nf-core/samtools/idxstats/** diff --git a/modules/nf-core/samtools/stats/environment.yml b/modules/nf-core/samtools/stats/environment.yml new file mode 100644 index 0000000..ed4e896 --- /dev/null +++ b/modules/nf-core/samtools/stats/environment.yml @@ -0,0 +1,7 @@ +name: samtools_stats +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/stats/main.nf b/modules/nf-core/samtools/stats/main.nf new file mode 100644 index 0000000..07286ef --- /dev/null +++ b/modules/nf-core/samtools/stats/main.nf @@ -0,0 +1,49 @@ +process SAMTOOLS_STATS { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : + 'biocontainers/samtools:1.17--h00cdaf9_0' }" + + input: + tuple val(meta), path(input), path(input_index) + tuple val(meta2), path(fasta) + + output: + tuple val(meta), path("*.stats"), emit: stats + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def reference = fasta ? "--reference ${fasta}" : "" + """ + samtools \\ + stats \\ + --threads ${task.cpus} \\ + ${reference} \\ + ${input} \\ + > ${prefix}.stats + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch ${prefix}.stats + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/stats/meta.yml b/modules/nf-core/samtools/stats/meta.yml new file mode 100644 index 0000000..735ff81 --- /dev/null +++ b/modules/nf-core/samtools/stats/meta.yml @@ -0,0 +1,63 @@ +name: samtools_stats +description: Produces comprehensive statistics from SAM/BAM/CRAM file +keywords: + - statistics + - counts + - bam + - sam + - cram +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - input: + type: file + description: BAM/CRAM file from alignment + pattern: "*.{bam,cram}" + - input_index: + type: file + description: BAI/CRAI file from alignment + pattern: "*.{bai,crai}" + - meta2: + type: map + description: | + Groovy Map containing reference information + e.g. [ id:'genome' ] + - fasta: + type: file + description: Reference file the CRAM was created with (optional) + pattern: "*.{fasta,fa}" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - stats: + type: file + description: File containing samtools stats output + pattern: "*.{stats}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@FriederikeHanssen" + - "@ramprasadn" +maintainers: + - "@drpatelh" + - "@FriederikeHanssen" + - "@ramprasadn" diff --git a/modules/nf-core/samtools/stats/tests/main.nf.test b/modules/nf-core/samtools/stats/tests/main.nf.test new file mode 100644 index 0000000..e037132 --- /dev/null +++ b/modules/nf-core/samtools/stats/tests/main.nf.test @@ -0,0 +1,78 @@ +nextflow_process { + + name "Test Process SAMTOOLS_STATS" + script "../main.nf" + process "SAMTOOLS_STATS" + tag "modules" + tag "modules/nf-core" + tag "samtools" + tag "samtools/stats" + + test("SAMTOOLS STATS Should run without failures") { + + when { + params { + + outdir = "$outputDir" + } + process { + """ + // define inputs of the process here. + input[0] = [ + [ id:'test', single_end:false ], // meta map + file(params.test_data['sarscov2']['illumina']['test_paired_end_sorted_bam'], checkIfExists: true), + file(params.test_data['sarscov2']['illumina']['test_paired_end_sorted_bam_bai'], checkIfExists: true) + + ] + input[1] = [[],[]] + """ + + } + } + + then { + assertAll( + {assert process.success}, + {assert snapshot(process.out).match()} + ) + } + + } + + test("SAMTOOLS CRAM Should run without failures") { + + when { + params { + + outdir = "$outputDir" + } + process { + """ + // define inputs of the process here + input[0] = [ + [ id:'test', single_end:false ], // meta map + file(params.test_data['homo_sapiens']['illumina']['test_paired_end_recalibrated_sorted_cram'], checkIfExists: true), + file(params.test_data['homo_sapiens']['illumina']['test_paired_end_recalibrated_sorted_cram_crai'], checkIfExists: true) + + ] + input[1] = [ + [ id:'genome' ], + file(params.test_data['homo_sapiens']['genome']['genome_fasta'], checkIfExists: true) + ] + """ + } + + + } + + then { + assertAll( + {assert process.success}, + {assert snapshot(process.out).match()} + ) + } + + } + + +} diff --git a/modules/nf-core/samtools/stats/tests/main.nf.test.snap b/modules/nf-core/samtools/stats/tests/main.nf.test.snap new file mode 100644 index 0000000..516b2b0 --- /dev/null +++ b/modules/nf-core/samtools/stats/tests/main.nf.test.snap @@ -0,0 +1,64 @@ +{ + "SAMTOOLS STATS Should run without failures": { + "content": [ + { + "0": [ + [ + { + "id": "test", + "single_end": false + }, + "test.stats:md5,6e768486d5df0257351c5419a79f9c9b" + ] + ], + "1": [ + "versions.yml:md5,08035f3409d934d47a416150884bb0df" + ], + "stats": [ + [ + { + "id": "test", + "single_end": false + }, + "test.stats:md5,6e768486d5df0257351c5419a79f9c9b" + ] + ], + "versions": [ + "versions.yml:md5,08035f3409d934d47a416150884bb0df" + ] + } + ], + "timestamp": "2023-10-18T12:12:42.998746" + }, + "SAMTOOLS CRAM Should run without failures": { + "content": [ + { + "0": [ + [ + { + "id": "test", + "single_end": false + }, + "test.stats:md5,7c9ee5747793cceb9d6f4d733345641a" + ] + ], + "1": [ + "versions.yml:md5,08035f3409d934d47a416150884bb0df" + ], + "stats": [ + [ + { + "id": "test", + "single_end": false + }, + "test.stats:md5,7c9ee5747793cceb9d6f4d733345641a" + ] + ], + "versions": [ + "versions.yml:md5,08035f3409d934d47a416150884bb0df" + ] + } + ], + "timestamp": "2023-10-18T12:13:30.747222" + } +} \ No newline at end of file diff --git a/modules/nf-core/samtools/stats/tests/tags.yml b/modules/nf-core/samtools/stats/tests/tags.yml new file mode 100644 index 0000000..7c28e30 --- /dev/null +++ b/modules/nf-core/samtools/stats/tests/tags.yml @@ -0,0 +1,2 @@ +samtools/stats: + - modules/nf-core/samtools/stats/** diff --git a/subworkflows/local/align_pacbio.nf b/subworkflows/local/align_pacbio.nf new file mode 100644 index 0000000..767563b --- /dev/null +++ b/subworkflows/local/align_pacbio.nf @@ -0,0 +1,70 @@ +// +// Align PacBio read files against the genome +// + +include { FILTER_PACBIO } from '../../subworkflows/local/filter_pacbio' +include { MINIMAP2_ALIGN } from '../../modules/nf-core/minimap2/align/main' +include { SAMTOOLS_MERGE } from '../../modules/nf-core/samtools/merge/main' +include { SAMTOOLS_SORT } from '../../modules/nf-core/samtools/sort/main' +include { CONVERT_STATS } from '../../subworkflows/local/convert_stats' + + +workflow ALIGN_PACBIO { + take: + fasta // channel: [ val(meta), /path/to/fasta ] + reads // channel: [ val(meta), /path/to/datafile ] + db // channel: /path/to/vector_db + + + main: + ch_versions = Channel.empty() + + + // Filter BAM and output as FASTQ + FILTER_PACBIO ( reads, db ) + ch_versions = ch_versions.mix ( FILTER_PACBIO.out.versions ) + + + // Align Fastq to Genome + fasta + | map { meta, file -> file } + | set { ch_fasta } + + MINIMAP2_ALIGN ( FILTER_PACBIO.out.fastq, ch_fasta, true, false, false ) + ch_versions = ch_versions.mix ( MINIMAP2_ALIGN.out.versions.first() ) + + + // Collect all alignment output by sample name + MINIMAP2_ALIGN.out.bam + | map { meta, bam -> [['id': meta.id.split('_')[0..-2].join('_'), 'datatype': meta.datatype], bam] } + | groupTuple ( by: [0] ) + | set { ch_bams } + + + // Merge + SAMTOOLS_MERGE ( ch_bams, [], [] ) + ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions.first() ) + + + // Position sort BAM file + SAMTOOLS_SORT ( SAMTOOLS_MERGE.out.bam ) + ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions.first() ) + + + // Convert merged BAM to CRAM and calculate indices and statistics + SAMTOOLS_SORT.out.bam + | map { meta, bam -> [ meta, bam, [] ] } + | set { ch_sort } + + CONVERT_STATS ( ch_sort, ch_fasta ) + ch_versions = ch_versions.mix ( CONVERT_STATS.out.versions ) + + + emit: + cram = CONVERT_STATS.out.cram // channel: [ val(meta), /path/to/cram ] + crai = CONVERT_STATS.out.crai // channel: [ val(meta), /path/to/crai ] + stats = CONVERT_STATS.out.stats // channel: [ val(meta), /path/to/stats ] + idxstats = CONVERT_STATS.out.idxstats // channel: [ val(meta), /path/to/idxstats ] + flagstat = CONVERT_STATS.out.flagstat // channel: [ val(meta), /path/to/flagstat ] + versions = ch_versions // channel: [ versions.yml ] +} diff --git a/subworkflows/local/convert_stats.nf b/subworkflows/local/convert_stats.nf new file mode 100644 index 0000000..7c381fd --- /dev/null +++ b/subworkflows/local/convert_stats.nf @@ -0,0 +1,54 @@ +// +// Convert BAM to CRAM, create index and calculate statistics +// + +include { SAMTOOLS_VIEW } from '../../modules/nf-core/samtools/view/main' +include { SAMTOOLS_STATS } from '../../modules/nf-core/samtools/stats/main' +include { SAMTOOLS_FLAGSTAT } from '../../modules/nf-core/samtools/flagstat/main' +include { SAMTOOLS_IDXSTATS } from '../../modules/nf-core/samtools/idxstats/main' + + +workflow CONVERT_STATS { + take: + bam // channel: [ val(meta), /path/to/bam, /path/to/bai] + fasta // channel: /path/to/fasta + + + main: + ch_versions = Channel.empty() + + + // Convert BAM to CRAM + SAMTOOLS_VIEW ( bam, fasta, [] ) + ch_versions = ch_versions.mix ( SAMTOOLS_VIEW.out.versions.first() ) + + + // Combine CRAM and CRAI into one channel + SAMTOOLS_VIEW.out.cram + | join ( SAMTOOLS_VIEW.out.crai ) + | set { ch_cram_crai } + + + // Calculate statistics + SAMTOOLS_STATS ( ch_cram_crai, fasta ) + ch_versions = ch_versions.mix ( SAMTOOLS_STATS.out.versions.first() ) + + + // Calculate statistics based on flag values + SAMTOOLS_FLAGSTAT ( ch_cram_crai ) + ch_versions = ch_versions.mix ( SAMTOOLS_FLAGSTAT.out.versions.first() ) + + + // Calculate index statistics + SAMTOOLS_IDXSTATS ( ch_cram_crai ) + ch_versions = ch_versions.mix ( SAMTOOLS_IDXSTATS.out.versions.first() ) + + + emit: + cram = SAMTOOLS_VIEW.out.cram // channel: [ val(meta), /path/to/cram ] + crai = SAMTOOLS_VIEW.out.crai // channel: [ val(meta), /path/to/crai ] + stats = SAMTOOLS_STATS.out.stats // channel: [ val(meta), /path/to/stats ] + flagstat = SAMTOOLS_FLAGSTAT.out.flagstat // channel: [ val(meta), /path/to/idxstats ] + idxstats = SAMTOOLS_IDXSTATS.out.idxstats // channel: [ val(meta), /path/to/flagstat ] + versions = ch_versions // channel: [ versions.yml ] +} diff --git a/subworkflows/local/filter_pacbio.nf b/subworkflows/local/filter_pacbio.nf new file mode 100644 index 0000000..6078d4d --- /dev/null +++ b/subworkflows/local/filter_pacbio.nf @@ -0,0 +1,77 @@ +// +// Filter PacBio reads +// Original protocol is a modified version by Shane of the original program, HiFiAdapterFilt +// + +include { SAMTOOLS_VIEW as SAMTOOLS_CONVERT } from '../../modules/nf-core/samtools/view/main' +include { SAMTOOLS_COLLATE } from '../../modules/nf-core/samtools/collate/main' +include { SAMTOOLS_FASTA } from '../../modules/nf-core/samtools/fasta/main' +include { GUNZIP } from '../../modules/nf-core/gunzip/main' +include { BLAST_BLASTN } from '../../modules/nf-core/blast/blastn/main' +include { PACBIO_FILTER } from '../../modules/local/pacbio_filter' +include { SAMTOOLS_VIEW as SAMTOOLS_FILTER } from '../../modules/nf-core/samtools/view/main' +include { SAMTOOLS_FASTQ } from '../../modules/nf-core/samtools/fastq/main' + + +workflow FILTER_PACBIO { + take: + reads // channel: [ val(meta), /path/to/datafile ] + db // channel: /path/to/vector_db + + + main: + ch_versions = Channel.empty() + + + // Convert from PacBio BAM to Samtools BAM + reads + | map { meta, bam -> [ meta, bam, [] ] } + | set { ch_pacbio } + + SAMTOOLS_CONVERT ( ch_pacbio, [], [] ) + ch_versions = ch_versions.mix ( SAMTOOLS_CONVERT.out.versions.first() ) + + + // Collate BAM file to create interleaved FASTA + SAMTOOLS_COLLATE ( SAMTOOLS_CONVERT.out.bam, [] ) + ch_versions = ch_versions.mix ( SAMTOOLS_COLLATE.out.versions.first() ) + + + // Convert BAM to FASTA + SAMTOOLS_FASTA ( SAMTOOLS_COLLATE.out.bam, true ) + ch_versions = ch_versions.mix ( SAMTOOLS_FASTA.out.versions.first() ) + + + // Gunzip FASTA file to BLAST + GUNZIP ( SAMTOOLS_FASTA.out.other ) + ch_versions = ch_versions.mix ( GUNZIP.out.versions.first() ) + + + // Nucleotide BLAST + BLAST_BLASTN ( GUNZIP.out.gunzip, db ) + ch_versions = ch_versions.mix ( BLAST_BLASTN.out.versions.first() ) + + + // Filter BLAST output + PACBIO_FILTER ( BLAST_BLASTN.out.txt ) + ch_versions = ch_versions.mix ( PACBIO_FILTER.out.versions.first() ) + + + // Create filtered BAM file + SAMTOOLS_CONVERT.out.bam + | join ( SAMTOOLS_CONVERT.out.csi ) + | set { ch_reads } + + SAMTOOLS_FILTER ( ch_reads, [], PACBIO_FILTER.out.list ) + ch_versions = ch_versions.mix ( SAMTOOLS_FILTER.out.versions.first() ) + + + // Convert BAM to FASTQ + SAMTOOLS_FASTQ ( SAMTOOLS_FILTER.out.unoutput, true ) + ch_versions = ch_versions.mix ( SAMTOOLS_FASTQ.out.versions.first() ) + + + emit: + fastq = SAMTOOLS_FASTQ.out.other // channel: [ meta, /path/to/fastq ] + versions = ch_versions // channel: [ versions.yml ] +} From a5b1d968e92fe68f891dcacb1770e79274fa550e Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 20 Nov 2023 10:28:59 +0000 Subject: [PATCH 076/123] add align option for pacbio_align subworkflow --- assets/vectorDB.tar.gz | Bin 0 -> 4640 bytes modules.json | 11 + .../nf-core/samtools/fastq/environment.yml | 7 + modules/nf-core/samtools/fastq/main.nf | 44 ++ modules/nf-core/samtools/fastq/meta.yml | 62 +++ modules/nf-core/untar/environment.yml | 9 + modules/nf-core/untar/main.nf | 63 +++ modules/nf-core/untar/meta.yml | 46 ++ modules/nf-core/untar/tests/main.nf.test | 77 +++ modules/nf-core/untar/tests/main.nf.test.snap | 513 ++++++++++++++++++ modules/nf-core/untar/tests/tags.yml | 2 + nextflow.config | 2 + nextflow_schema.json | 10 + workflows/variantcalling.nf | 30 +- 14 files changed, 875 insertions(+), 1 deletion(-) create mode 100644 assets/vectorDB.tar.gz create mode 100644 modules/nf-core/samtools/fastq/environment.yml create mode 100644 modules/nf-core/samtools/fastq/main.nf create mode 100644 modules/nf-core/samtools/fastq/meta.yml create mode 100644 modules/nf-core/untar/environment.yml create mode 100644 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meta.single_end ? "-1 ${prefix}_1.fastq.gz -s ${prefix}_singleton.fastq.gz" : + "-1 ${prefix}_1.fastq.gz -2 ${prefix}_2.fastq.gz -s ${prefix}_singleton.fastq.gz" + """ + samtools \\ + fastq \\ + $args \\ + --threads ${task.cpus-1} \\ + -0 ${prefix}_other.fastq.gz \\ + $input \\ + $output + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/samtools/fastq/meta.yml b/modules/nf-core/samtools/fastq/meta.yml new file mode 100644 index 0000000..c4002a4 --- /dev/null +++ b/modules/nf-core/samtools/fastq/meta.yml @@ -0,0 +1,62 @@ +name: samtools_fastq +description: Converts a SAM/BAM/CRAM file to FASTQ +keywords: + - bam + - sam + - cram + - fastq +tools: + - samtools: + description: | + SAMtools is a set of utilities for interacting with and post-processing + short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. + These files are generated as output by short read aligners like BWA. + homepage: http://www.htslib.org/ + documentation: http://www.htslib.org/doc/samtools.html + doi: 10.1093/bioinformatics/btp352 + licence: ["MIT"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - input: + type: file + description: BAM/CRAM/SAM file + pattern: "*.{bam,cram,sam}" + - interleave: + type: boolean + description: Set true for interleaved fastq file +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + - fastq: + type: file + description: Compressed FASTQ file(s) with reads with either the READ1 or READ2 flag set in separate files. + pattern: "*_{1,2}.fastq.gz" + - interleaved: + type: file + description: Compressed FASTQ file with reads with either the READ1 or READ2 flag set in a combined file. Needs collated input file. + pattern: "*_interleaved.fastq.gz" + - singleton: + type: file + description: Compressed FASTQ file with singleton reads + pattern: "*_singleton.fastq.gz" + - other: + type: file + description: Compressed FASTQ file with reads with either both READ1 and READ2 flags set or unset + pattern: "*_other.fastq.gz" +authors: + - "@priyanka-surana" + - "@suzannejin" +maintainers: + - "@priyanka-surana" + - "@suzannejin" diff --git a/modules/nf-core/untar/environment.yml b/modules/nf-core/untar/environment.yml new file mode 100644 index 0000000..d6917da --- /dev/null +++ b/modules/nf-core/untar/environment.yml @@ -0,0 +1,9 @@ +name: untar +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - conda-forge::sed=4.7 + - conda-forge::grep=3.11 + - conda-forge::tar=1.34 diff --git a/modules/nf-core/untar/main.nf b/modules/nf-core/untar/main.nf new file mode 100644 index 0000000..8a75bb9 --- /dev/null +++ b/modules/nf-core/untar/main.nf @@ -0,0 +1,63 @@ +process UNTAR { + tag "$archive" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/ubuntu:20.04' : + 'nf-core/ubuntu:20.04' }" + + input: + tuple val(meta), path(archive) + + output: + tuple val(meta), path("$prefix"), emit: untar + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def args2 = task.ext.args2 ?: '' + prefix = task.ext.prefix ?: ( meta.id ? "${meta.id}" : archive.baseName.toString().replaceFirst(/\.tar$/, "")) + + """ + mkdir $prefix + + ## Ensures --strip-components only applied when top level of tar contents is a directory + ## If just files or multiple directories, place all in prefix + if [[ \$(tar -taf ${archive} | grep -o -P "^.*?\\/" | uniq | wc -l) -eq 1 ]]; then + tar \\ + -C $prefix --strip-components 1 \\ + -xavf \\ + $args \\ + $archive \\ + $args2 + else + tar \\ + -C $prefix \\ + -xavf \\ + $args \\ + $archive \\ + $args2 + fi + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + untar: \$(echo \$(tar --version 2>&1) | sed 's/^.*(GNU tar) //; s/ Copyright.*\$//') + END_VERSIONS + """ + + stub: + prefix = task.ext.prefix ?: ( meta.id ? "${meta.id}" : archive.toString().replaceFirst(/\.[^\.]+(.gz)?$/, "")) + """ + mkdir $prefix + touch ${prefix}/file.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + untar: \$(echo \$(tar --version 2>&1) | sed 's/^.*(GNU tar) //; s/ Copyright.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/untar/meta.yml b/modules/nf-core/untar/meta.yml new file mode 100644 index 0000000..a9a2110 --- /dev/null +++ b/modules/nf-core/untar/meta.yml @@ -0,0 +1,46 @@ +name: untar +description: Extract files. +keywords: + - untar + - uncompress + - extract +tools: + - untar: + description: | + Extract tar.gz files. + documentation: https://www.gnu.org/software/tar/manual/ + licence: ["GPL-3.0-or-later"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - archive: + type: file + description: File to be untar + pattern: "*.{tar}.{gz}" +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - untar: + type: directory + description: Directory containing contents of archive + pattern: "*/" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" + - "@matthdsm" + - "@jfy133" +maintainers: + - "@joseespinosa" + - "@drpatelh" + - "@matthdsm" + - "@jfy133" diff --git a/modules/nf-core/untar/tests/main.nf.test b/modules/nf-core/untar/tests/main.nf.test new file mode 100644 index 0000000..d40db13 --- /dev/null +++ b/modules/nf-core/untar/tests/main.nf.test @@ -0,0 +1,77 @@ +nextflow_process { + + name "Test Process UNTAR" + script "../main.nf" + process "UNTAR" + + tag "modules" + tag "modules_nfcore" + tag "untar" + + test("test_untar") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ [], file(params.test_data['sarscov2']['genome']['kraken2_tar_gz'], checkIfExists: true) ] + """ + } + } + + then { + assertAll ( + { assert process.success }, + { assert snapshot(process.out.untar).match("test_untar") }, + ) + } + + } + + test("test_untar_different_output_path") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ [], file(params.test_data['homo_sapiens']['illumina']['test_flowcell'], checkIfExists: true) ] + """ + } + } + + then { + assertAll ( + { assert process.success }, + { assert snapshot(process.out.untar).match("test_untar_different_output_path") }, + ) + } + + } + + test("test_untar_onlyfiles") { + + when { + params { + outdir = "$outputDir" + } + process { + """ + input[0] = [ [], file(params.test_data['generic']['tar']['tar_gz'], checkIfExists: true) ] + """ + } + } + + then { + assertAll ( + { assert process.success }, + { assert snapshot(process.out.untar).match("test_untar_onlyfiles") }, + ) + } + + } + +} diff --git a/modules/nf-core/untar/tests/main.nf.test.snap b/modules/nf-core/untar/tests/main.nf.test.snap new file 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+ }, + "test_untar": { + "content": [ + [ + [ + [ + + ], + [ + "hash.k2d:md5,8b8598468f54a7087c203ad0190555d9", + "opts.k2d:md5,a033d00cf6759407010b21700938f543", + "taxo.k2d:md5,094d5891cdccf2f1468088855c214b2c" + ] + ] + ] + ], + "timestamp": "2023-10-18T11:56:08.16574" + } +} \ No newline at end of file diff --git a/modules/nf-core/untar/tests/tags.yml b/modules/nf-core/untar/tests/tags.yml new file mode 100644 index 0000000..feb6f15 --- /dev/null +++ b/modules/nf-core/untar/tests/tags.yml @@ -0,0 +1,2 @@ +untar: + - modules/nf-core/untar/** diff --git a/nextflow.config b/nextflow.config index 90da5d8..bcc78b0 100644 --- a/nextflow.config +++ b/nextflow.config @@ -13,10 +13,12 @@ params { input = null fasta = null fai = null + align = false interval = null include_positions = null exclude_positions = null split_fasta_cutoff = 100000 + vector_db = "${projectDir}/assets/vectorDB.tar.gz" // Boilerplate options outdir = 'results' diff --git a/nextflow_schema.json b/nextflow_schema.json index e035219..958cdb9 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -28,6 +28,12 @@ "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.", "fa_icon": "fas fa-folder-open" }, + "vector_db": { + "type": "string", + "default": "/Users/gq2/git3/variantcalling/assets/vectorDB.tar.gz", + "description": "Path to directory or tar.gz archive for pre-built PacBio vector database.", + "format": "file-path" + }, "email": { "type": "string", "description": "Email address for completion summary.", @@ -51,6 +57,10 @@ "type": "string", "description": "Path to the index file of the FASTA genome file, either fai or gzi." }, + "align": { + "type": "boolean", + "description": "Align the input reads to the reference" + }, "interval": { "type": "string", "description": "Interval bed file." diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index c88480f..54f442f 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -60,11 +60,13 @@ if ( (params.include_positions) && (params.exclude_positions) ){ // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // include { INPUT_CHECK } from '../subworkflows/local/input_check' +include { ALIGN_PACBIO } from '../subworkflows/local/align_pacbio' include { INPUT_MERGE } from '../subworkflows/local/input_merge' include { INPUT_FILTER_SPLIT } from '../subworkflows/local/input_filter_split' include { DEEPVARIANT_CALLER } from '../subworkflows/local/deepvariant_caller' include { PROCESS_VCF } from '../subworkflows/local/process_vcf' + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT NF-CORE MODULES/SUBWORKFLOWS @@ -75,7 +77,8 @@ include { PROCESS_VCF } from '../subworkflows/local/process_vcf' // MODULE: Installed directly from nf-core/modules // include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' -include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' +include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' +include { UNTAR } from '../modules/nf-core/untar/main' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -127,6 +130,31 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix( INPUT_CHECK.out.versions ) + // + // SUBWORKFLOW: align reads if required + // + if( params.align ){ + + if ( params.vector_db.endsWith( '.tar.gz' ) ) { + + UNTAR ( [ [:], params.vector_db ] ).untar + | map { meta, file -> file } + | set { ch_vector_db } + ch_versions = ch_versions.mix ( UNTAR.out.versions ) + } else { + Channel.fromPath ( params.vector_db ) + | set { ch_vector_db } + } + + ALIGN_PACBIO ( + ch_fasta, + INPUT_CHECK.out.reads, + ch_vector_db + ) + ch_versions = ch_versions.mix( ALIGN_PACBIO.out.versions ) + } + + // // SUBWORKFLOW: merge the input reads by sample name // From 4c8367e60879412b7ee2ae66c3ac0a32576ebd15 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 23 Nov 2023 11:02:57 +0000 Subject: [PATCH 077/123] re-patch samtools view --- modules.json | 2 +- modules/nf-core/samtools/view/environment.yml | 7 ++++++ modules/nf-core/samtools/view/main.nf | 5 +++-- modules/nf-core/samtools/view/meta.yml | 5 +++++ .../nf-core/samtools/view/samtools-view.diff | 22 +++++++++++++++++++ 5 files changed, 38 insertions(+), 3 deletions(-) create mode 100644 modules/nf-core/samtools/view/environment.yml create mode 100644 modules/nf-core/samtools/view/samtools-view.diff diff --git a/modules.json b/modules.json index 6ef43e2..bc30640 100644 --- a/modules.json +++ b/modules.json @@ -88,7 +88,7 @@ }, "samtools/view": { "branch": "master", - "git_sha": "3ffae3598260a99e8db3207dead9f73f87f90d1f", + "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", "installed_by": ["modules"], "patch": "modules/nf-core/samtools/view/samtools-view.diff" }, diff --git a/modules/nf-core/samtools/view/environment.yml b/modules/nf-core/samtools/view/environment.yml new file mode 100644 index 0000000..141e7bd --- /dev/null +++ b/modules/nf-core/samtools/view/environment.yml @@ -0,0 +1,7 @@ +name: samtools_view +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::samtools=1.17 diff --git a/modules/nf-core/samtools/view/main.nf b/modules/nf-core/samtools/view/main.nf index cb91fac..613c6e7 100644 --- a/modules/nf-core/samtools/view/main.nf +++ b/modules/nf-core/samtools/view/main.nf @@ -2,7 +2,7 @@ process SAMTOOLS_VIEW { tag "$meta.id" label 'process_low' - conda "bioconda::samtools=1.17" + conda "${moduleDir}/environment.yml" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : 'biocontainers/samtools:1.17--h00cdaf9_0' }" @@ -19,6 +19,7 @@ process SAMTOOLS_VIEW { tuple val(meta), path("*.bai"), emit: bai, optional: true tuple val(meta), path("*.csi"), emit: csi, optional: true tuple val(meta), path("*.crai"), emit: crai, optional: true + tuple val(meta), path("*.unoutput"), emit: unoutput, optional: true path "versions.yml", emit: versions when: @@ -29,7 +30,7 @@ process SAMTOOLS_VIEW { def args2 = task.ext.args2 ?: '' def prefix = task.ext.prefix ?: "${meta.id}" def reference = fasta ? "--reference ${fasta}" : "" - def readnames = qname ? "--qname-file ${qname}": "" + def readnames = qname ? "--qname-file ${qname} --unoutput ${prefix}.unoutput": "" def file_type = args.contains("--output-fmt sam") ? "sam" : args.contains("--output-fmt bam") ? "bam" : args.contains("--output-fmt cram") ? "cram" : diff --git a/modules/nf-core/samtools/view/meta.yml b/modules/nf-core/samtools/view/meta.yml index 3b05450..3dadafa 100644 --- a/modules/nf-core/samtools/view/meta.yml +++ b/modules/nf-core/samtools/view/meta.yml @@ -82,3 +82,8 @@ authors: - "@joseespinosa" - "@FriederikeHanssen" - "@priyanka-surana" +maintainers: + - "@drpatelh" + - "@joseespinosa" + - "@FriederikeHanssen" + - "@priyanka-surana" diff --git a/modules/nf-core/samtools/view/samtools-view.diff b/modules/nf-core/samtools/view/samtools-view.diff new file mode 100644 index 0000000..1fa860a --- /dev/null +++ b/modules/nf-core/samtools/view/samtools-view.diff @@ -0,0 +1,22 @@ +Changes in module 'nf-core/samtools/view' +--- modules/nf-core/samtools/view/main.nf ++++ modules/nf-core/samtools/view/main.nf +@@ -19,6 +19,7 @@ + tuple val(meta), path("*.bai"), emit: bai, optional: true + tuple val(meta), path("*.csi"), emit: csi, optional: true + tuple val(meta), path("*.crai"), emit: crai, optional: true ++ tuple val(meta), path("*.unoutput"), emit: unoutput, optional: true + path "versions.yml", emit: versions + + when: +@@ -29,7 +30,7 @@ + def args2 = task.ext.args2 ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def reference = fasta ? "--reference ${fasta}" : "" +- def readnames = qname ? "--qname-file ${qname}": "" ++ def readnames = qname ? "--qname-file ${qname} --unoutput ${prefix}.unoutput": "" + def file_type = args.contains("--output-fmt sam") ? "sam" : + args.contains("--output-fmt bam") ? "bam" : + args.contains("--output-fmt cram") ? "cram" : + +************************************************************ From b4f582dc924b07de09ab2bf0421fd82e075b47c2 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 27 Nov 2023 10:08:56 +0000 Subject: [PATCH 078/123] nf-core modules update vcftools --- modules.json | 2 +- modules/nf-core/vcftools/environment.yml | 1 + modules/nf-core/vcftools/main.nf | 1 - modules/nf-core/vcftools/vcftools.diff | 33 ------------------------ 4 files changed, 2 insertions(+), 35 deletions(-) delete mode 100644 modules/nf-core/vcftools/vcftools.diff diff --git a/modules.json b/modules.json index bc30640..e7bcfa1 100644 --- a/modules.json +++ b/modules.json @@ -99,7 +99,7 @@ }, "vcftools": { "branch": "master", - "git_sha": "8fc1d24c710ebe1d5de0f2447ec9439fd3d9d66a", + "git_sha": "485558b40040fc3ace093d9084210125d8ba4c97", "installed_by": ["modules"], "patch": "modules/nf-core/vcftools/vcftools.diff" } diff --git a/modules/nf-core/vcftools/environment.yml b/modules/nf-core/vcftools/environment.yml index 875817e..503449e 100644 --- a/modules/nf-core/vcftools/environment.yml +++ b/modules/nf-core/vcftools/environment.yml @@ -1,3 +1,4 @@ +name: vcftools channels: - conda-forge - bioconda diff --git a/modules/nf-core/vcftools/main.nf b/modules/nf-core/vcftools/main.nf index b36c429..0e61955 100644 --- a/modules/nf-core/vcftools/main.nf +++ b/modules/nf-core/vcftools/main.nf @@ -94,7 +94,6 @@ process VCFTOOLS { (args.contains('--hapcount')) ? "--hapcount ${bed}" : (args.contains('--positions')) ? "--positions ${bed}" : (args.contains('--exclude-positions')) ? "--exclude-positions ${bed}" : '' - args_list.removeIf { it.contains('--bed') } args_list.removeIf { it.contains('--exclude-bed') } args_list.removeIf { it.contains('--hapcount') } diff --git a/modules/nf-core/vcftools/vcftools.diff b/modules/nf-core/vcftools/vcftools.diff deleted file mode 100644 index fd2e9ec..0000000 --- a/modules/nf-core/vcftools/vcftools.diff +++ /dev/null @@ -1,33 +0,0 @@ -Changes in module 'nf-core/vcftools' ---- modules/nf-core/vcftools/meta.yml -+++ modules/nf-core/vcftools/meta.yml -@@ -1,6 +1,7 @@ - name: vcftools - description: A set of tools written in Perl and C++ for working with VCF files - keywords: -+ - VCFtools - - VCF - - sort - tools: - ---- modules/nf-core/vcftools/main.nf -+++ modules/nf-core/vcftools/main.nf -@@ -91,10 +91,15 @@ - - def bed_arg = (args.contains('--bed')) ? "--bed ${bed}" : - (args.contains('--exclude-bed')) ? "--exclude-bed ${bed}" : -- (args.contains('--hapcount')) ? "--hapcount ${bed}" : '' -+ (args.contains('--hapcount')) ? "--hapcount ${bed}" : -+ (args.contains('--positions')) ? "--positions ${bed}" : -+ (args.contains('--exclude-positions')) ? "--exclude-positions ${bed}" : '' -+ - args_list.removeIf { it.contains('--bed') } - args_list.removeIf { it.contains('--exclude-bed') } - args_list.removeIf { it.contains('--hapcount') } -+ args_list.removeIf { it.contains('--positions') } -+ args_list.removeIf { it.contains('--exclude-positions') } - - def diff_variant_arg = (args.contains('--diff')) ? "--diff ${diff_variant_file}" : - (args.contains('--gzdiff')) ? "--gzdiff ${diff_variant_file}" : - -************************************************************ From f2cefafc13a71f71a679431a478916b5efc4d8c6 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 27 Nov 2023 16:07:12 +0000 Subject: [PATCH 079/123] update the way calling module or subworkflow --- bin/pacbio_filter.sh | 6 ++++ conf/modules.config | 46 +++++++++++++++++++++++++++++ subworkflows/local/filter_pacbio.nf | 7 +++-- workflows/variantcalling.nf | 23 +++++++-------- 4 files changed, 67 insertions(+), 15 deletions(-) create mode 100755 bin/pacbio_filter.sh diff --git a/bin/pacbio_filter.sh b/bin/pacbio_filter.sh new file mode 100755 index 0000000..73d7caa --- /dev/null +++ b/bin/pacbio_filter.sh @@ -0,0 +1,6 @@ +#!/bin/bash + +input=$1 +output=$2 + +grep -v 'MG551957' $input | awk -v OFS='\t' '{if (($2 ~ /NGB00972/ && $3 >= 97 && $4 >= 44) || ($2 ~ /NGB00973/ && $3 >= 97 && $4 >= 34) || ($2 ~ /^bc/ && $3 >= 99 && $4 >= 16)) print $1}' | sort -u > $output diff --git a/conf/modules.config b/conf/modules.config index cb326e7..c8bbc2f 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -20,6 +20,52 @@ process { ] } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_CONVERT' { + ext.args = "-be '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index" + } + + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FILTER' { + ext.prefix = { "${meta.id}.filter" } + } + + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_COLLATE' { + ext.prefix = { "${meta.id}.collate" } + } + + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:BLAST_BLASTN' { + ext.args = '-task blastn -reward 1 -penalty -5 -gapopen 3 -gapextend 3 -dust yes -soft_masking true -evalue .01 -searchsp 1750000000000 -outfmt 6' + } + + withName: '.*:.*:ALIGN_PACBIO:MINIMAP2_ALIGN' { + ext.args = '-ax map-hifi --cs=short' + } + + withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_MERGE' { + ext.args = { "-c -p" } + ext.prefix = { "${meta.id}.merge" } + } + + withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_SORT' { + ext.prefix = { "${meta.id}.sort" } + } + + withName: '.*:CONVERT_STATS:SAMTOOLS_VIEW' { + ext.prefix = { "${fasta.baseName}.${meta.datatype}.${meta.id}" } + ext.args = '--output-fmt cram --write-index' + } + + withName: '.*:CONVERT_STATS:SAMTOOLS_STATS' { + ext.prefix = { "${input.baseName}" } + } + + withName: '.*:CONVERT_STATS:SAMTOOLS_FLAGSTAT' { + ext.prefix = { "${bam.baseName}" } + } + + withName: '.*:CONVERT_STATS:SAMTOOLS_IDXSTATS' { + ext.prefix = { "${bam.baseName}" } + } + withName: '.*:INPUT_FILTER_SPLIT:SAMTOOLS_VIEW' { ext.args = '--output-fmt cram --write-index -F 0x900' ext.prefix = { "${meta.id}_filtered" } diff --git a/subworkflows/local/filter_pacbio.nf b/subworkflows/local/filter_pacbio.nf index 6078d4d..2e306bf 100644 --- a/subworkflows/local/filter_pacbio.nf +++ b/subworkflows/local/filter_pacbio.nf @@ -28,7 +28,7 @@ workflow FILTER_PACBIO { | map { meta, bam -> [ meta, bam, [] ] } | set { ch_pacbio } - SAMTOOLS_CONVERT ( ch_pacbio, [], [] ) + SAMTOOLS_CONVERT (ch_pacbio, [ [], [] ], [] ) ch_versions = ch_versions.mix ( SAMTOOLS_CONVERT.out.versions.first() ) @@ -48,7 +48,8 @@ workflow FILTER_PACBIO { // Nucleotide BLAST - BLAST_BLASTN ( GUNZIP.out.gunzip, db ) + db.map{db -> [ [], db]}.set{ch_db} + BLAST_BLASTN ( GUNZIP.out.gunzip, ch_db ) ch_versions = ch_versions.mix ( BLAST_BLASTN.out.versions.first() ) @@ -62,7 +63,7 @@ workflow FILTER_PACBIO { | join ( SAMTOOLS_CONVERT.out.csi ) | set { ch_reads } - SAMTOOLS_FILTER ( ch_reads, [], PACBIO_FILTER.out.list ) + SAMTOOLS_FILTER ( ch_reads, [ [], [] ], PACBIO_FILTER.out.list ) ch_versions = ch_versions.mix ( SAMTOOLS_FILTER.out.versions.first() ) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 54f442f..b200086 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -44,12 +44,6 @@ if ( (params.include_positions) && (params.exclude_positions) ){ ch_positions = [] } -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - CONFIG FILES -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT LOCAL MODULES/SUBWORKFLOWS @@ -66,7 +60,6 @@ include { INPUT_FILTER_SPLIT } from '../subworkflows/local/input_filter_split' include { DEEPVARIANT_CALLER } from '../subworkflows/local/deepvariant_caller' include { PROCESS_VCF } from '../subworkflows/local/process_vcf' - /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT NF-CORE MODULES/SUBWORKFLOWS @@ -122,6 +115,7 @@ workflow VARIANTCALLING { ch_index = ch_fai } + // // SUBWORKFLOW: Read in samplesheet, validate and stage input files // @@ -130,6 +124,7 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix( INPUT_CHECK.out.versions ) + // // SUBWORKFLOW: align reads if required // @@ -146,11 +141,12 @@ workflow VARIANTCALLING { | set { ch_vector_db } } - ALIGN_PACBIO ( - ch_fasta, - INPUT_CHECK.out.reads, - ch_vector_db - ) + ch_fasta.map{ fasta -> [[], fasta]}.set{fasta_meta} + ALIGN_PACBIO ( + fasta_meta, + INPUT_CHECK.out.reads, + ch_vector_db + ) ch_versions = ch_versions.mix( ALIGN_PACBIO.out.versions ) } @@ -177,6 +173,7 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix( INPUT_FILTER_SPLIT.out.versions ) + // // SUBWORKFLOW: call deepvariant // @@ -185,6 +182,7 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix( DEEPVARIANT_CALLER.out.versions ) + // // convert VCF channel meta id // @@ -198,6 +196,7 @@ workflow VARIANTCALLING { PROCESS_VCF( vcf, ch_positions ) ch_versions = ch_versions.mix( PROCESS_VCF.out.versions ) + // // MODULE: Combine different version together // From 27e5d9e318259ddef15b8fe5bd359fbfec7a915f Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 27 Nov 2023 16:07:41 +0000 Subject: [PATCH 080/123] add a new test_align profile --- assets/samplesheet_test_align.csv | 2 ++ conf/test_align.config | 27 +++++++++++++++++++++++++++ nextflow.config | 1 + 3 files changed, 30 insertions(+) create mode 100644 assets/samplesheet_test_align.csv create mode 100644 conf/test_align.config diff --git a/assets/samplesheet_test_align.csv b/assets/samplesheet_test_align.csv new file mode 100644 index 0000000..381e21d --- /dev/null +++ b/assets/samplesheet_test_align.csv @@ -0,0 +1,2 @@ +sample,datatype,datafile +mMelMel3,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Meles_meles/genomic_data/mMelMel3/hic/35528_2%231.subset.cram \ No newline at end of file diff --git a/conf/test_align.config b/conf/test_align.config new file mode 100644 index 0000000..9680e67 --- /dev/null +++ b/conf/test_align.config @@ -0,0 +1,27 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Nextflow config file for running minimal tests +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Defines input files and everything required to run a fast and simple pipeline test. + + Use as follows: + nextflow run sanger-tol/variantcalling -profile test, --outdir + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Test align profile' + config_profile_description = 'Minimal test dataset to check pipeline function' + + // Limit resources so that this can run on GitHub Actions + max_cpus = 2 + max_memory = '6.GB' + max_time = '6.h' + + // Input data + input = "${projectDir}/assets/samplesheet_test_align.csv" + + // Fasta references + fasta = "https://tolit.cog.sanger.ac.uk/test-data/Meles_meles/assembly/release/mMelMel3.1_paternal_haplotype/GCA_922984935.2.subset.fasta" +} diff --git a/nextflow.config b/nextflow.config index bcc78b0..93512e2 100644 --- a/nextflow.config +++ b/nextflow.config @@ -169,6 +169,7 @@ profiles { executor.memory = 60.GB } test { includeConfig 'conf/test.config' } + test_align { includeConfig 'conf/test_align.config' } test_full { includeConfig 'conf/test_full.config' } } From 59892b0203a7d3836666a707217fcb7b4d0e8df7 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 27 Nov 2023 16:43:07 +0000 Subject: [PATCH 081/123] make samtools convert output as bam --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index c8bbc2f..29fb899 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -21,7 +21,7 @@ process { } withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_CONVERT' { - ext.args = "-be '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index" + ext.args = "-be '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index --output-fmt bam" } withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FILTER' { From eef4d82093a870087847627f69bdb63a5b8b28c2 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 28 Nov 2023 09:46:30 +0000 Subject: [PATCH 082/123] use compressed fasta file --- conf/test_align.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_align.config b/conf/test_align.config index 9680e67..746d4a8 100644 --- a/conf/test_align.config +++ b/conf/test_align.config @@ -23,5 +23,5 @@ params { input = "${projectDir}/assets/samplesheet_test_align.csv" // Fasta references - fasta = "https://tolit.cog.sanger.ac.uk/test-data/Meles_meles/assembly/release/mMelMel3.1_paternal_haplotype/GCA_922984935.2.subset.fasta" + fasta = "https://tolit.cog.sanger.ac.uk/test-data/Meles_meles/assembly/release/mMelMel3.1_paternal_haplotype/GCA_922984935.2.subset.fasta.gz" } From 9347f34508247f87e2761fc530c373ae3eeadc07 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 28 Nov 2023 10:51:13 +0000 Subject: [PATCH 083/123] extra modules configs --- conf/modules.config | 14 +++++++++----- 1 file changed, 9 insertions(+), 5 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 29fb899..f909984 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -24,10 +24,6 @@ process { ext.args = "-be '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index --output-fmt bam" } - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FILTER' { - ext.prefix = { "${meta.id}.filter" } - } - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_COLLATE' { ext.prefix = { "${meta.id}.collate" } } @@ -36,8 +32,16 @@ process { ext.args = '-task blastn -reward 1 -penalty -5 -gapopen 3 -gapextend 3 -dust yes -soft_masking true -evalue .01 -searchsp 1750000000000 -outfmt 6' } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FILTER' { + ext.prefix = { "${meta.id}.filter" } + } + + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FASTQ' { + ext.args = '-F 0x200 -nt' + } + withName: '.*:.*:ALIGN_PACBIO:MINIMAP2_ALIGN' { - ext.args = '-ax map-hifi --cs=short' + ext.args = { "-ax map-hifi --cs=short -R ${meta.read_group}" } } withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_MERGE' { From 2d70af5fed6ff976350cef3e58e43cad3147cb8e Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 28 Nov 2023 18:01:39 +0000 Subject: [PATCH 084/123] remove -b flag for samtools view --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index f909984..a92a73d 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -21,7 +21,7 @@ process { } withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_CONVERT' { - ext.args = "-be '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index --output-fmt bam" + ext.args = "-e '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index --output-fmt bam" } withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_COLLATE' { From f7d5faf742155f3d866aa7b62dd557ad67f1e171 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 11:02:47 +0000 Subject: [PATCH 085/123] add RG group to meta data in sample checking --- subworkflows/local/deepvariant_caller.nf | 6 ++--- subworkflows/local/input_check.nf | 30 ++++++++++++++++++++---- subworkflows/local/input_merge.nf | 2 +- 3 files changed, 29 insertions(+), 9 deletions(-) diff --git a/subworkflows/local/deepvariant_caller.nf b/subworkflows/local/deepvariant_caller.nf index 10b5359..31b42e0 100644 --- a/subworkflows/local/deepvariant_caller.nf +++ b/subworkflows/local/deepvariant_caller.nf @@ -14,7 +14,7 @@ workflow DEEPVARIANT_CALLER { ch_versions = Channel.empty() reads_fasta.map { meta, cram, crai, interval, fasta_file_name, fasta, fai -> - [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.type ], + [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.datatype ], cram, crai, interval @@ -23,14 +23,14 @@ workflow DEEPVARIANT_CALLER { // fasta fasta = reads_fasta.map { meta, cram, crai, interval, fasta_file_name, fasta, fai -> - [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.type ], + [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.datatype ], fasta ] } // fai fai = reads_fasta.map{ meta, cram, crai, interval, fasta_file_name, fasta, fai -> - [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.type ], + [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.datatype ], fai ] } diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index 7e9f667..ec8a9f4 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -12,13 +12,33 @@ workflow INPUT_CHECK { SAMPLESHEET_CHECK ( samplesheet ) .csv .splitCsv ( header:true, sep:',' ) - .map { [ - [ id: it.sample, sample: it.sample, type: it.datatype ], - file(it.datafile) - ] } + .map { create_data_channel( it ) } .set { reads } - + emit: reads // channel: [ val(meta), data ] versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ] } + +// Function to get list of [ meta, reads ] +def create_data_channel ( LinkedHashMap row ) { + // create meta map + def meta = [:] + meta.id = row.sample + meta.sample = row.sample + meta.datatype = row.datatype + + if ( meta.datatype == "pacbio" ) { + platform = "PACBIO" + } + meta.read_group = "\'@RG\\tID:" + row.datafile.split('/')[-1].split('\\.')[0] + "\\tPL:" + platform + "\\tSM:" + meta.sample + "\'" + + // add path(s) of the read file(s) to the meta map + def data_meta = [] + if ( !file(row.datafile).exists() ) { + exit 1, "ERROR: Please check input samplesheet -> Data file does not exist!\n${row.datafile}" + } else { + data_meta = [ meta, file(row.datafile) ] + } + return data_meta +} diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 90bb82f..c63c679 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -38,7 +38,7 @@ workflow INPUT_MERGE { .join( grouped_reads ) .map { sample, meta, bam_cram_list -> [ [ id: ( bam_cram_list.size() == 1 ) ? sample : sample + '_combined', - type: meta.type + type: meta.datatype ], bam_cram_list ]} From ce47ad9c0cb61b11133fc01476c66b2f79cd9c6b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 11:38:30 +0000 Subject: [PATCH 086/123] update channel with meta --- subworkflows/local/align_pacbio.nf | 11 ++++------- subworkflows/local/convert_stats.nf | 5 ++--- 2 files changed, 6 insertions(+), 10 deletions(-) diff --git a/subworkflows/local/align_pacbio.nf b/subworkflows/local/align_pacbio.nf index 767563b..5c31f34 100644 --- a/subworkflows/local/align_pacbio.nf +++ b/subworkflows/local/align_pacbio.nf @@ -26,23 +26,20 @@ workflow ALIGN_PACBIO { // Align Fastq to Genome - fasta - | map { meta, file -> file } - | set { ch_fasta } - MINIMAP2_ALIGN ( FILTER_PACBIO.out.fastq, ch_fasta, true, false, false ) + MINIMAP2_ALIGN ( FILTER_PACBIO.out.fastq, fasta, true, false, false ) ch_versions = ch_versions.mix ( MINIMAP2_ALIGN.out.versions.first() ) // Collect all alignment output by sample name MINIMAP2_ALIGN.out.bam - | map { meta, bam -> [['id': meta.id.split('_')[0..-2].join('_'), 'datatype': meta.datatype], bam] } + | map { meta, bam -> [['id': meta.id, 'datatype': meta.datatype], bam] } | groupTuple ( by: [0] ) | set { ch_bams } // Merge - SAMTOOLS_MERGE ( ch_bams, [], [] ) + SAMTOOLS_MERGE ( ch_bams, [ [], [] ], [ [], [] ] ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions.first() ) @@ -56,7 +53,7 @@ workflow ALIGN_PACBIO { | map { meta, bam -> [ meta, bam, [] ] } | set { ch_sort } - CONVERT_STATS ( ch_sort, ch_fasta ) + CONVERT_STATS ( ch_sort, fasta ) ch_versions = ch_versions.mix ( CONVERT_STATS.out.versions ) diff --git a/subworkflows/local/convert_stats.nf b/subworkflows/local/convert_stats.nf index 7c381fd..9118e8d 100644 --- a/subworkflows/local/convert_stats.nf +++ b/subworkflows/local/convert_stats.nf @@ -11,15 +11,14 @@ include { SAMTOOLS_IDXSTATS } from '../../modules/nf-core/samtools/idxstats/main workflow CONVERT_STATS { take: bam // channel: [ val(meta), /path/to/bam, /path/to/bai] - fasta // channel: /path/to/fasta + fasta // channel: [ val(meta), /path/to/fasta ] main: ch_versions = Channel.empty() - // Convert BAM to CRAM - SAMTOOLS_VIEW ( bam, fasta, [] ) + SAMTOOLS_VIEW ( bam, fasta, [ ] ) ch_versions = ch_versions.mix ( SAMTOOLS_VIEW.out.versions.first() ) From 80e83c78c98dfd3a94ae4de6ee3561c1113ea339 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 11:50:08 +0000 Subject: [PATCH 087/123] publish aligned cram files with stats --- conf/modules.config | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index a92a73d..2ce358a 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -70,6 +70,14 @@ process { ext.prefix = { "${bam.baseName}" } } + withName: '.*:ALIGN_PACBIO:CONVERT_STATS:.*' { + publishDir = [ + path: { "${params.outdir}/variant_calling" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + withName: '.*:INPUT_FILTER_SPLIT:SAMTOOLS_VIEW' { ext.args = '--output-fmt cram --write-index -F 0x900' ext.prefix = { "${meta.id}_filtered" } From 8e2ccc8cf052df815951f253ede12b02ff1396d0 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 13:33:37 +0000 Subject: [PATCH 088/123] Remove samtools sort after minimap_align and before merging. --- subworkflows/local/align_pacbio.nf | 11 ++--------- 1 file changed, 2 insertions(+), 9 deletions(-) diff --git a/subworkflows/local/align_pacbio.nf b/subworkflows/local/align_pacbio.nf index 5c31f34..26c575a 100644 --- a/subworkflows/local/align_pacbio.nf +++ b/subworkflows/local/align_pacbio.nf @@ -26,30 +26,23 @@ workflow ALIGN_PACBIO { // Align Fastq to Genome - MINIMAP2_ALIGN ( FILTER_PACBIO.out.fastq, fasta, true, false, false ) ch_versions = ch_versions.mix ( MINIMAP2_ALIGN.out.versions.first() ) // Collect all alignment output by sample name MINIMAP2_ALIGN.out.bam - | map { meta, bam -> [['id': meta.id, 'datatype': meta.datatype], bam] } + | map { meta, bam -> [['id': meta.id, 'datatype': meta.datatype, 'sample': meta.sample ], bam] } | groupTuple ( by: [0] ) | set { ch_bams } - // Merge SAMTOOLS_MERGE ( ch_bams, [ [], [] ], [ [], [] ] ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions.first() ) - // Position sort BAM file - SAMTOOLS_SORT ( SAMTOOLS_MERGE.out.bam ) - ch_versions = ch_versions.mix ( SAMTOOLS_SORT.out.versions.first() ) - - // Convert merged BAM to CRAM and calculate indices and statistics - SAMTOOLS_SORT.out.bam + SAMTOOLS_MERGE.out.bam | map { meta, bam -> [ meta, bam, [] ] } | set { ch_sort } From 2815232aee217faf7111449dd8cfc2070797b0e0 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 14:39:07 +0000 Subject: [PATCH 089/123] add conditions pacbio align modules configs --- conf/modules.config | 91 +++++++++++++++++++++++---------------------- 1 file changed, 47 insertions(+), 44 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 2ce358a..a2f44ea 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -20,62 +20,65 @@ process { ] } - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_CONVERT' { - ext.args = "-e '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index --output-fmt bam" - } + if( params.align ) { - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_COLLATE' { - ext.prefix = { "${meta.id}.collate" } - } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_CONVERT' { + ext.args = "-e '[rq]>=0.99' -x fi -x fp -x ri -x rp --write-index --output-fmt bam" + } - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:BLAST_BLASTN' { - ext.args = '-task blastn -reward 1 -penalty -5 -gapopen 3 -gapextend 3 -dust yes -soft_masking true -evalue .01 -searchsp 1750000000000 -outfmt 6' - } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_COLLATE' { + ext.prefix = { "${meta.id}.collate" } + } - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FILTER' { - ext.prefix = { "${meta.id}.filter" } - } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:BLAST_BLASTN' { + ext.args = '-task blastn -reward 1 -penalty -5 -gapopen 3 -gapextend 3 -dust yes -soft_masking true -evalue .01 -searchsp 1750000000000 -outfmt 6' + } - withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FASTQ' { - ext.args = '-F 0x200 -nt' - } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FILTER' { + ext.prefix = { "${meta.id}.filter" } + } - withName: '.*:.*:ALIGN_PACBIO:MINIMAP2_ALIGN' { - ext.args = { "-ax map-hifi --cs=short -R ${meta.read_group}" } - } + withName: '.*:ALIGN_PACBIO:FILTER_PACBIO:SAMTOOLS_FASTQ' { + ext.args = '-F 0x200 -nt' + } - withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_MERGE' { - ext.args = { "-c -p" } - ext.prefix = { "${meta.id}.merge" } - } + withName: '.*:.*:ALIGN_PACBIO:MINIMAP2_ALIGN' { + ext.args = { "-ax map-hifi --cs=short -R ${meta.read_group}" } + } - withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_SORT' { - ext.prefix = { "${meta.id}.sort" } - } + withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_MERGE' { + ext.args = { "-c -p" } + ext.prefix = { "${meta.id}.merge" } + } - withName: '.*:CONVERT_STATS:SAMTOOLS_VIEW' { - ext.prefix = { "${fasta.baseName}.${meta.datatype}.${meta.id}" } - ext.args = '--output-fmt cram --write-index' - } + withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_SORT' { + ext.prefix = { "${meta.id}.sort" } + } - withName: '.*:CONVERT_STATS:SAMTOOLS_STATS' { - ext.prefix = { "${input.baseName}" } - } + withName: '.*:CONVERT_STATS:SAMTOOLS_VIEW' { + ext.prefix = { "${fasta.baseName}.${meta.datatype}.${meta.id}" } + ext.args = '--output-fmt cram --write-index' + } - withName: '.*:CONVERT_STATS:SAMTOOLS_FLAGSTAT' { - ext.prefix = { "${bam.baseName}" } - } + withName: '.*:CONVERT_STATS:SAMTOOLS_STATS' { + ext.prefix = { "${input.baseName}" } + } - withName: '.*:CONVERT_STATS:SAMTOOLS_IDXSTATS' { - ext.prefix = { "${bam.baseName}" } - } + withName: '.*:CONVERT_STATS:SAMTOOLS_FLAGSTAT' { + ext.prefix = { "${bam.baseName}" } + } - withName: '.*:ALIGN_PACBIO:CONVERT_STATS:.*' { - publishDir = [ - path: { "${params.outdir}/variant_calling" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] + withName: '.*:CONVERT_STATS:SAMTOOLS_IDXSTATS' { + ext.prefix = { "${bam.baseName}" } + } + + withName: '.*:ALIGN_PACBIO:CONVERT_STATS:.*' { + publishDir = [ + path: { "${params.outdir}/variant_calling" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } } withName: '.*:INPUT_FILTER_SPLIT:SAMTOOLS_VIEW' { From 7125e18e39271c92f9d7f679e69f0b5d4439f1b5 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 14:40:01 +0000 Subject: [PATCH 090/123] pass the aligned reads to variant calling --- workflows/variantcalling.nf | 11 ++++++++--- 1 file changed, 8 insertions(+), 3 deletions(-) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index b200086..714cdc3 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -141,15 +141,20 @@ workflow VARIANTCALLING { | set { ch_vector_db } } - ch_fasta.map{ fasta -> [[], fasta]}.set{fasta_meta} + ch_fasta.map{ fasta -> [[], fasta] }.set{ fasta_meta } ALIGN_PACBIO ( fasta_meta, INPUT_CHECK.out.reads, ch_vector_db ) ch_versions = ch_versions.mix( ALIGN_PACBIO.out.versions ) - } + ch_aligned_reads = ALIGN_PACBIO.out.cram + + } else { + + ch_aligned_reads = INPUT_CHECK.out.reads + } // // SUBWORKFLOW: merge the input reads by sample name @@ -157,7 +162,7 @@ workflow VARIANTCALLING { INPUT_MERGE ( ch_fasta, ch_index, - INPUT_CHECK.out.reads, + ch_aligned_reads, ) ch_versions = ch_versions.mix( INPUT_MERGE.out.versions ) From adcda19b2a4835d0a6e26f4e3e3d15946727257b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 16:18:55 +0000 Subject: [PATCH 091/123] remove unused module config --- assets/samplesheet_test_align.csv | 2 +- conf/modules.config | 4 ---- conf/test_align.config | 2 +- 3 files changed, 2 insertions(+), 6 deletions(-) diff --git a/assets/samplesheet_test_align.csv b/assets/samplesheet_test_align.csv index 381e21d..a2bae25 100644 --- a/assets/samplesheet_test_align.csv +++ b/assets/samplesheet_test_align.csv @@ -1,2 +1,2 @@ sample,datatype,datafile -mMelMel3,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Meles_meles/genomic_data/mMelMel3/hic/35528_2%231.subset.cram \ No newline at end of file +icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/genomic_data/icCanRufa1/pacbio/m64094_200730_174533.ccs.bc1010_BAK8A_OA--bc1010_BAK8A_OA_0_03.bam diff --git a/conf/modules.config b/conf/modules.config index a2f44ea..7a0528e 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -51,10 +51,6 @@ process { ext.prefix = { "${meta.id}.merge" } } - withName: '.*:.*:ALIGN_PACBIO:SAMTOOLS_SORT' { - ext.prefix = { "${meta.id}.sort" } - } - withName: '.*:CONVERT_STATS:SAMTOOLS_VIEW' { ext.prefix = { "${fasta.baseName}.${meta.datatype}.${meta.id}" } ext.args = '--output-fmt cram --write-index' diff --git a/conf/test_align.config b/conf/test_align.config index 746d4a8..6284ff5 100644 --- a/conf/test_align.config +++ b/conf/test_align.config @@ -23,5 +23,5 @@ params { input = "${projectDir}/assets/samplesheet_test_align.csv" // Fasta references - fasta = "https://tolit.cog.sanger.ac.uk/test-data/Meles_meles/assembly/release/mMelMel3.1_paternal_haplotype/GCA_922984935.2.subset.fasta.gz" + fasta = "https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/assembly/GCA_947369205.1_OX376310.1_CANBKR010000003.1.fasta.gz" } From 6b545450ec8d0352d4e922516de3809652c638a7 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 17:17:49 +0000 Subject: [PATCH 092/123] no need to sort merge after aligment --- conf/modules.config | 9 +++++---- subworkflows/local/input_merge.nf | 2 +- workflows/variantcalling.nf | 29 +++++++++++++++-------------- 3 files changed, 21 insertions(+), 19 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 7a0528e..e963a11 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -75,6 +75,11 @@ process { saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] } + } else{ + + withName: '.*:INPUT_MERGE:SAMTOOLS_MERGE' { + ext.args = '--write-index' + } } withName: '.*:INPUT_FILTER_SPLIT:SAMTOOLS_VIEW' { @@ -82,10 +87,6 @@ process { ext.prefix = { "${meta.id}_filtered" } } - withName: '.*:INPUT_MERGE:SAMTOOLS_MERGE' { - ext.args = '--write-index' - } - withName: '.*:DEEPVARIANT_CALLER:DEEPVARIANT' { ext.args = '--model_type=PACBIO' } diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index c63c679..bbbec3e 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -38,7 +38,7 @@ workflow INPUT_MERGE { .join( grouped_reads ) .map { sample, meta, bam_cram_list -> [ [ id: ( bam_cram_list.size() == 1 ) ? sample : sample + '_combined', - type: meta.datatype + datatype: meta.datatype ], bam_cram_list ]} diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 714cdc3..50970c5 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -149,30 +149,31 @@ workflow VARIANTCALLING { ) ch_versions = ch_versions.mix( ALIGN_PACBIO.out.versions ) - ch_aligned_reads = ALIGN_PACBIO.out.cram + ALIGN_PACBIO.out.cram + .join( ALIGN_PACBIO.out.crai ) + .set{ ch_aligned_reads } } else { - ch_aligned_reads = INPUT_CHECK.out.reads - } - - // - // SUBWORKFLOW: merge the input reads by sample name - // - INPUT_MERGE ( - ch_fasta, - ch_index, - ch_aligned_reads, - ) - ch_versions = ch_versions.mix( INPUT_MERGE.out.versions ) + // + // SUBWORKFLOW: merge the input reads by sample name + // + INPUT_MERGE ( + ch_fasta, + ch_index, + INPUT_CHECK.out.reads, + ) + ch_versions = ch_versions.mix( INPUT_MERGE.out.versions ) + ch_aligned_reads = INPUT_MERGE.out.indexed_merged_reads + } // // SUBWORKFLOW: split the input fasta file and filter input reads // INPUT_FILTER_SPLIT ( ch_fasta, - INPUT_MERGE.out.indexed_merged_reads, + ch_aligned_reads, ch_interval, split_fasta_cutoff ) From e03a4ee5f38a25ff8ee20a32e327b5491d125f6f Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Wed, 29 Nov 2023 17:48:35 +0000 Subject: [PATCH 093/123] add two test sample files for alignments --- assets/samplesheet_test_align.csv | 1 + 1 file changed, 1 insertion(+) diff --git a/assets/samplesheet_test_align.csv b/assets/samplesheet_test_align.csv index a2bae25..b7f429e 100644 --- a/assets/samplesheet_test_align.csv +++ b/assets/samplesheet_test_align.csv @@ -1,2 +1,3 @@ sample,datatype,datafile icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/genomic_data/icCanRufa1/pacbio/m64094_200730_174533.ccs.bc1010_BAK8A_OA--bc1010_BAK8A_OA_0_03.bam +icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/genomic_data/icCanRufa1/pacbio/m64094_200730_174533.ccs.bc1010_BAK8A_OA--bc1010_BAK8A_OA_0_02.bam From 1540017f25dd8e590281518c3825514bde316de4 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 11:32:52 +0000 Subject: [PATCH 094/123] convert fasta channel to value channel --- subworkflows/local/align_pacbio.nf | 1 + subworkflows/local/input_merge.nf | 11 ++++----- workflows/variantcalling.nf | 37 ++++++++++++++---------------- 3 files changed, 22 insertions(+), 27 deletions(-) diff --git a/subworkflows/local/align_pacbio.nf b/subworkflows/local/align_pacbio.nf index 26c575a..12a8255 100644 --- a/subworkflows/local/align_pacbio.nf +++ b/subworkflows/local/align_pacbio.nf @@ -36,6 +36,7 @@ workflow ALIGN_PACBIO { | groupTuple ( by: [0] ) | set { ch_bams } + // Merge SAMTOOLS_MERGE ( ch_bams, [ [], [] ], [ [], [] ] ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions.first() ) diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index bbbec3e..4df8957 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -7,8 +7,8 @@ include { SAMTOOLS_SORT } from '../../modules/nf-core/samtools/sort' workflow INPUT_MERGE { take: - fasta // file: /path/to/genome.fasta or /path/to/genome.fasta.gz - fai // file: /path/to/genome.*.fai or /path/to/genome.fasta.gz.gzi + fasta // channel: [ val(meta), /path/to/genome.fasta or /path/to/genome.fasta.gz ] + fai // channel: [ val(meta), /path/to/genome.*.fai or /path/to/genome.fasta.gz.gzi ] reads // channel: [ val(meta), data ] main: @@ -45,12 +45,9 @@ workflow INPUT_MERGE { .set { grouped_reads_with_meta } // call samtool merge - ch_fasta = fasta.map { fasta -> [ [ 'id': fasta.baseName ], fasta ] }.first() - ch_fai = fai.map { fai -> [ [ 'id': fai.baseName ], fai ] }.first() - SAMTOOLS_MERGE( grouped_reads_with_meta, - ch_fasta, - ch_fai + fasta, + fai ) ch_versions = ch_versions.mix ( SAMTOOLS_MERGE.out.versions ) diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index 50970c5..ef6cde9 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -84,38 +84,32 @@ include { UNTAR } from '../modules/nf-core/untar/main' workflow VARIANTCALLING { ch_versions = Channel.empty() + ch_fasta + .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } + .first() + .set { ch_genome } // // check reference fasta index given or not // if( params.fai == null ){ - ch_fasta - .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } - .set { ch_genome } - SAMTOOLS_FAIDX ( ch_genome, [[], []] ) ch_versions = ch_versions.mix( SAMTOOLS_FAIDX.out.versions ) - SAMTOOLS_FAIDX.out.fai - .map{ mata, fai -> fai } - .set{ ch_fai } - - SAMTOOLS_FAIDX.out.gzi - .map{ meta, gzi -> gzi } - .set{ ch_gzi } - if( params.fasta.endsWith('.gz') ){ - ch_index = ch_gzi + ch_genome_index = SAMTOOLS_FAIDX.out.gzi }else{ - ch_index = ch_fai + ch_genome_index = SAMTOOLS_FAIDX.out.fai } }else{ - ch_index = ch_fai + ch_index + .map { fai -> [ [ 'id': fai.baseName ], fai ] } + .first() + .set { ch_genome_index } } - // // SUBWORKFLOW: Read in samplesheet, validate and stage input files // @@ -136,14 +130,17 @@ workflow VARIANTCALLING { | map { meta, file -> file } | set { ch_vector_db } ch_versions = ch_versions.mix ( UNTAR.out.versions ) + + } else { + Channel.fromPath ( params.vector_db ) | set { ch_vector_db } + } - ch_fasta.map{ fasta -> [[], fasta] }.set{ fasta_meta } ALIGN_PACBIO ( - fasta_meta, + ch_genome, INPUT_CHECK.out.reads, ch_vector_db ) @@ -159,8 +156,8 @@ workflow VARIANTCALLING { // SUBWORKFLOW: merge the input reads by sample name // INPUT_MERGE ( - ch_fasta, - ch_index, + ch_genome, + ch_genome_index, INPUT_CHECK.out.reads, ) ch_versions = ch_versions.mix( INPUT_MERGE.out.versions ) From 8463d43733dd750708e55652f4d868b9cc3fd57d Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 11:58:50 +0000 Subject: [PATCH 095/123] put _T1 back to distinguish the same samles --- bin/check_samplesheet.py | 6 +++++- subworkflows/local/input_check.nf | 4 ++-- 2 files changed, 7 insertions(+), 3 deletions(-) diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py index d088e65..c91f170 100755 --- a/bin/check_samplesheet.py +++ b/bin/check_samplesheet.py @@ -108,7 +108,11 @@ def validate_unique_samples(self): """ if len(self._seen) != len(self.validated): raise AssertionError("The combination of sample name and data file must be unique.") - + seen = Counter() + for row in self.validated: + sample = row[self._sample_col] + seen[sample] += 1 + row[self._sample_col] = f"{sample}_T{seen[sample]}" def read_head(handle, num_lines=10): """Read the specified number of lines from the current position in the file.""" diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf index ec8a9f4..b71f3fd 100644 --- a/subworkflows/local/input_check.nf +++ b/subworkflows/local/input_check.nf @@ -25,13 +25,13 @@ def create_data_channel ( LinkedHashMap row ) { // create meta map def meta = [:] meta.id = row.sample - meta.sample = row.sample + meta.sample = row.sample.split('_')[0..-2].join('_') meta.datatype = row.datatype if ( meta.datatype == "pacbio" ) { platform = "PACBIO" } - meta.read_group = "\'@RG\\tID:" + row.datafile.split('/')[-1].split('\\.')[0] + "\\tPL:" + platform + "\\tSM:" + meta.sample + "\'" + meta.read_group = "\'@RG\\tID:" + row.datafile.split('/')[-1].split('\\.')[0..-2].join('.') + "\\tPL:" + platform + "\\tSM:" + meta.sample + "\'" // add path(s) of the read file(s) to the meta map def data_meta = [] From 50df394e0ec07a98ff9ea6f37ab64a893b95dd6c Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 12:15:55 +0000 Subject: [PATCH 096/123] add combined in the aligned bam/cram file name if sample being combined --- assets/samplesheet_test_align.csv | 1 + subworkflows/local/align_pacbio.nf | 14 ++++++++++++-- 2 files changed, 13 insertions(+), 2 deletions(-) diff --git a/assets/samplesheet_test_align.csv b/assets/samplesheet_test_align.csv index b7f429e..4b5a9b2 100644 --- a/assets/samplesheet_test_align.csv +++ b/assets/samplesheet_test_align.csv @@ -1,3 +1,4 @@ sample,datatype,datafile icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/genomic_data/icCanRufa1/pacbio/m64094_200730_174533.ccs.bc1010_BAK8A_OA--bc1010_BAK8A_OA_0_03.bam icCanRufa1,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/genomic_data/icCanRufa1/pacbio/m64094_200730_174533.ccs.bc1010_BAK8A_OA--bc1010_BAK8A_OA_0_02.bam +icCanRufa1XXXXX,pacbio,https://tolit.cog.sanger.ac.uk/test-data/Cantharis_rufa/genomic_data/icCanRufa1/pacbio/m64094_200730_174533.ccs.bc1010_BAK8A_OA--bc1010_BAK8A_OA_0_02.bam diff --git a/subworkflows/local/align_pacbio.nf b/subworkflows/local/align_pacbio.nf index 12a8255..4c98f2c 100644 --- a/subworkflows/local/align_pacbio.nf +++ b/subworkflows/local/align_pacbio.nf @@ -32,10 +32,20 @@ workflow ALIGN_PACBIO { // Collect all alignment output by sample name MINIMAP2_ALIGN.out.bam - | map { meta, bam -> [['id': meta.id, 'datatype': meta.datatype, 'sample': meta.sample ], bam] } + | map { meta, bam -> [['id': meta.sample, 'datatype': meta.datatype, 'sample': meta.sample ], bam] } | groupTuple ( by: [0] ) + | map { meta, bam_list -> + [ + [ + 'id': ( bam_list.size() == 1 ) ? meta.sample : meta.sample + '_combined', + 'sample' : meta.sample, + 'datatype': meta.datatype + ], + bam_list + ] + } | set { ch_bams } - + // Merge SAMTOOLS_MERGE ( ch_bams, [ [], [] ], [ [], [] ] ) From add2bcb53877eea227dbc8e061ef2d2bd299dcd4 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 12:26:31 +0000 Subject: [PATCH 097/123] add a full test with aligment --- assets/samplesheet_test_full_align.csv | 2 ++ conf/test_align.config | 6 +++--- conf/test_full_align.config | 25 +++++++++++++++++++++++++ nextflow.config | 7 ++++--- 4 files changed, 34 insertions(+), 6 deletions(-) create mode 100644 assets/samplesheet_test_full_align.csv create mode 100644 conf/test_full_align.config diff --git a/assets/samplesheet_test_full_align.csv b/assets/samplesheet_test_full_align.csv new file mode 100644 index 0000000..d5b0ac4 --- /dev/null +++ b/assets/samplesheet_test_full_align.csv @@ -0,0 +1,2 @@ +sample,datatype,datafile +ilPolIcar1,pacbio,/lustre/scratch124/tol/projects/darwin/data/insects/Polyommatus_icarus/genomic_data/ilPolIcar1/pacbio/m64016_191206_183623.ccs.bc1019_BAK8B_OA--bc1019_BAK8B_OA.bam diff --git a/conf/test_align.config b/conf/test_align.config index 6284ff5..8da6b65 100644 --- a/conf/test_align.config +++ b/conf/test_align.config @@ -5,14 +5,14 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run sanger-tol/variantcalling -profile test, --outdir + nextflow run sanger-tol/variantcalling -profile test_align, --outdir --align ---------------------------------------------------------------------------------------- */ params { - config_profile_name = 'Test align profile' - config_profile_description = 'Minimal test dataset to check pipeline function' + config_profile_name = 'Test profile with alignment' + config_profile_description = 'Minimal unaligned test dataset to check pipeline function' // Limit resources so that this can run on GitHub Actions max_cpus = 2 diff --git a/conf/test_full_align.config b/conf/test_full_align.config new file mode 100644 index 0000000..79b9fd7 --- /dev/null +++ b/conf/test_full_align.config @@ -0,0 +1,25 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Nextflow config file for running full-size tests with alignment +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Defines input files and everything required to run a full size pipeline test. + + Use as follows: + nextflow run sanger-tol/variantcalling -profile test_full_align, --outdir --align + +---------------------------------------------------------------------------------------- +*/ + +cleanup = true + +params { + config_profile_name = 'Full test profile with alignment' + config_profile_description = 'Full non-aligned test dataset to check pipeline function' + + // Input data for full size test + input = "${projectDir}/assets/samplesheet_test_full_align.csv" + + // Fasta references + fasta = "/lustre/scratch124/tol/projects/darwin/data/insects/Polyommatus_icarus/assembly/release/ilPolIcar1.1/insdc/GCA_937595015.1.fasta.gz" + +} diff --git a/nextflow.config b/nextflow.config index 93512e2..bb1f0f6 100644 --- a/nextflow.config +++ b/nextflow.config @@ -168,9 +168,10 @@ profiles { executor.cpus = 16 executor.memory = 60.GB } - test { includeConfig 'conf/test.config' } - test_align { includeConfig 'conf/test_align.config' } - test_full { includeConfig 'conf/test_full.config' } + test { includeConfig 'conf/test.config' } + test_align { includeConfig 'conf/test_align.config' } + test_full { includeConfig 'conf/test_full.config' } + test_full_align { includeConfig 'conf/test_full_align.config' } } From 4285d4c373bae0f69ba5907315aa88d8baf1c26e Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 12:34:48 +0000 Subject: [PATCH 098/123] add one alignment test in the simple ci test --- .github/workflows/ci.yml | 4 ++++ 1 file changed, 4 insertions(+) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index dbc6e02..567ac13 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -41,3 +41,7 @@ jobs: # Remember that you can parallelise this by using strategy.matrix run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results + + - name: Run pipeline with unaligned test data + run: | + nextflow run ${GITHUB_WORKSPACE} -profile test_align,docker --outdir ./results --align From ef82d4975e69d5da2be0a5e6a0efd730d1800736 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 12:38:23 +0000 Subject: [PATCH 099/123] black check --- bin/check_samplesheet.py | 1 + 1 file changed, 1 insertion(+) diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py index c91f170..3a6b9d7 100755 --- a/bin/check_samplesheet.py +++ b/bin/check_samplesheet.py @@ -114,6 +114,7 @@ def validate_unique_samples(self): seen[sample] += 1 row[self._sample_col] = f"{sample}_T{seen[sample]}" + def read_head(handle, num_lines=10): """Read the specified number of lines from the current position in the file.""" lines = [] From 6bf3a19daafbb6ee83cac92411384109359889f7 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 13:16:13 +0000 Subject: [PATCH 100/123] EditorConfig linting --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index e963a11..22d84ce 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -67,7 +67,7 @@ process { withName: '.*:CONVERT_STATS:SAMTOOLS_IDXSTATS' { ext.prefix = { "${bam.baseName}" } } - + withName: '.*:ALIGN_PACBIO:CONVERT_STATS:.*' { publishDir = [ path: { "${params.outdir}/variant_calling" }, From 006eced3bdb8450099226a67ef529f78ac78082b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 30 Nov 2023 13:25:22 +0000 Subject: [PATCH 101/123] make the sanger farm test with alignment --- .github/workflows/sanger_test.yml | 3 ++- .github/workflows/sanger_test_full.yml | 3 ++- 2 files changed, 4 insertions(+), 2 deletions(-) diff --git a/.github/workflows/sanger_test.yml b/.github/workflows/sanger_test.yml index 28f7625..a4e5426 100644 --- a/.github/workflows/sanger_test.yml +++ b/.github/workflows/sanger_test.yml @@ -19,8 +19,9 @@ jobs: parameters: | { "outdir": "${{ secrets.TOWER_WORKDIR_PARENT }}/results/${{ github.repository }}/results-${{ github.sha }}", + "align": true } - profiles: test,sanger,singularity,cleanup + profiles: test_align,sanger,singularity,cleanup - uses: actions/upload-artifact@v3 with: diff --git a/.github/workflows/sanger_test_full.yml b/.github/workflows/sanger_test_full.yml index e028c6b..a552c63 100644 --- a/.github/workflows/sanger_test_full.yml +++ b/.github/workflows/sanger_test_full.yml @@ -32,8 +32,9 @@ jobs: parameters: | { "outdir": "${{ secrets.TOWER_WORKDIR_PARENT }}/results/${{ github.repository }}/results-${{ env.REVISION }}", + "align": true, } - profiles: test_full,sanger,singularity,cleanup + profiles: test_full_align,sanger,singularity,cleanup - uses: actions/upload-artifact@v3 with: From 9a0003ae4fb4cabdf0d7ba14de89e89cd0edb4ba Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Fri, 1 Dec 2023 10:59:32 +0000 Subject: [PATCH 102/123] Update module 'samtools/fasta --- modules.json | 2 +- modules/nf-core/samtools/fasta/main.nf | 2 +- modules/nf-core/samtools/fasta/meta.yml | 2 +- 3 files changed, 3 insertions(+), 3 deletions(-) diff --git a/modules.json b/modules.json index e7bcfa1..f5561b1 100644 --- a/modules.json +++ b/modules.json @@ -52,7 +52,7 @@ }, "samtools/fasta": { "branch": "master", - "git_sha": "3f5420aa22e00bd030a2556dfdffc9e164ec0ec5", + "git_sha": "6f4299292ef2c5b66e6829527b2647c301b77cc9", "installed_by": ["modules"] }, "samtools/fastq": { diff --git a/modules/nf-core/samtools/fasta/main.nf b/modules/nf-core/samtools/fasta/main.nf index 925ed62..dc4ad98 100644 --- a/modules/nf-core/samtools/fasta/main.nf +++ b/modules/nf-core/samtools/fasta/main.nf @@ -38,7 +38,7 @@ process SAMTOOLS_FASTA { cat <<-END_VERSIONS > versions.yml "${task.process}": - samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//' )) + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') END_VERSIONS """ } diff --git a/modules/nf-core/samtools/fasta/meta.yml b/modules/nf-core/samtools/fasta/meta.yml index 1d07ea1..eae26f0 100644 --- a/modules/nf-core/samtools/fasta/meta.yml +++ b/modules/nf-core/samtools/fasta/meta.yml @@ -12,7 +12,7 @@ tools: documentation: "https://www.htslib.org/doc/samtools-fasta.html" tool_dev_url: "https://github.com/samtools/samtools" doi: "10.1093/bioinformatics/btp352" - licence: "['MIT']" + licence: ["MIT"] input: # Only when we have meta - meta: From 4db4cbc9c904c19d0a798c740ce60dc007cf3721 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 7 Dec 2023 10:50:17 +0000 Subject: [PATCH 103/123] remove the combined in the output file name if the same sample input file being combined. --- subworkflows/local/align_pacbio.nf | 10 ---------- subworkflows/local/input_merge.nf | 2 +- 2 files changed, 1 insertion(+), 11 deletions(-) diff --git a/subworkflows/local/align_pacbio.nf b/subworkflows/local/align_pacbio.nf index 4c98f2c..75f4ac2 100644 --- a/subworkflows/local/align_pacbio.nf +++ b/subworkflows/local/align_pacbio.nf @@ -34,16 +34,6 @@ workflow ALIGN_PACBIO { MINIMAP2_ALIGN.out.bam | map { meta, bam -> [['id': meta.sample, 'datatype': meta.datatype, 'sample': meta.sample ], bam] } | groupTuple ( by: [0] ) - | map { meta, bam_list -> - [ - [ - 'id': ( bam_list.size() == 1 ) ? meta.sample : meta.sample + '_combined', - 'sample' : meta.sample, - 'datatype': meta.datatype - ], - bam_list - ] - } | set { ch_bams } diff --git a/subworkflows/local/input_merge.nf b/subworkflows/local/input_merge.nf index 4df8957..12f2653 100644 --- a/subworkflows/local/input_merge.nf +++ b/subworkflows/local/input_merge.nf @@ -37,7 +37,7 @@ workflow INPUT_MERGE { .map { sample, meta_list -> [sample, meta_list[0]] } .join( grouped_reads ) .map { sample, meta, bam_cram_list -> [ - [ id: ( bam_cram_list.size() == 1 ) ? sample : sample + '_combined', + [ id: sample, datatype: meta.datatype ], bam_cram_list From 625f649545cdafdc2bc91dedf8a7edac8639dc4b Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 7 Dec 2023 13:47:40 +0000 Subject: [PATCH 104/123] update file name for VCf output files --- subworkflows/local/deepvariant_caller.nf | 26 +++++++++++++++++++----- 1 file changed, 21 insertions(+), 5 deletions(-) diff --git a/subworkflows/local/deepvariant_caller.nf b/subworkflows/local/deepvariant_caller.nf index 31b42e0..7e4adf0 100644 --- a/subworkflows/local/deepvariant_caller.nf +++ b/subworkflows/local/deepvariant_caller.nf @@ -14,7 +14,11 @@ workflow DEEPVARIANT_CALLER { ch_versions = Channel.empty() reads_fasta.map { meta, cram, crai, interval, fasta_file_name, fasta, fai -> - [ [ id: meta.id + "_" + fasta_file_name, sample: meta.id, type: meta.datatype ], + [ [ id: meta.id + "_" + fasta_file_name, + sample: meta.id, + type: meta.datatype, + fasta_file_name: fasta_file_name + ], cram, crai, interval @@ -44,9 +48,15 @@ workflow DEEPVARIANT_CALLER { // group the vcf files together by sample DEEPVARIANT.out.vcf - .map { meta, vcf -> [ meta.sample, vcf ] } + .map { meta, vcf -> [ + [ id: meta.fasta_file_name.tokenize(".")[0..-2].join(".") + + "." + meta.type + + "." + meta.sample + ], + vcf + ] } .groupTuple() - .map { sample, vcf -> [ [id: sample], vcf, [] ] } + .map { meta, vcf -> [ meta, vcf, [] ] } .set { vcf } // catcat vcf files @@ -55,9 +65,15 @@ workflow DEEPVARIANT_CALLER { // group the g vcf files together by sample DEEPVARIANT.out.gvcf - .map { meta, gvcf -> [ meta.sample, gvcf ] } + .map { meta, gvcf -> [ + [ id: meta.fasta_file_name.tokenize(".")[0..-2].join(".") + + "." + meta.type + + "." + meta.sample + ], + gvcf + ] } .groupTuple() - .map { sample, gvcf -> [ [ id: sample ], gvcf, [] ] } + .map { meta, gvcf -> [ meta, gvcf, [] ] } .set { g_vcf } // catcat g vcf files From d348591fec9c86414619a98d1836d30a4358528e Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 7 Dec 2023 15:33:59 +0000 Subject: [PATCH 105/123] make sure fasta or fa being removed from the fasta file name --- conf/modules.config | 2 +- workflows/variantcalling.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 22d84ce..acc63f5 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -52,7 +52,7 @@ process { } withName: '.*:CONVERT_STATS:SAMTOOLS_VIEW' { - ext.prefix = { "${fasta.baseName}.${meta.datatype}.${meta.id}" } + ext.prefix = { "${meta2.id}.${meta.datatype}.${meta.id}" } ext.args = '--output-fmt cram --write-index' } diff --git a/workflows/variantcalling.nf b/workflows/variantcalling.nf index ef6cde9..f3e7450 100644 --- a/workflows/variantcalling.nf +++ b/workflows/variantcalling.nf @@ -85,7 +85,7 @@ workflow VARIANTCALLING { ch_versions = Channel.empty() ch_fasta - .map { fasta -> [ [ 'id': fasta.baseName ], fasta ] } + .map { fasta -> [ [ 'id': fasta.baseName - '.fasta' - '.fa' ], fasta ] } .first() .set { ch_genome } From 7d20b1d2db04c13576dcd10ff5e37170f49d832f Mon Sep 17 00:00:00 2001 From: Guoying Qi <729395+gq1@users.noreply.github.com> Date: Thu, 7 Dec 2023 15:42:08 +0000 Subject: [PATCH 106/123] Update nextflow_schema.json wrong default value Co-authored-by: Matthieu Muffato --- nextflow_schema.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 958cdb9..c857174 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -30,7 +30,7 @@ }, "vector_db": { "type": "string", - "default": "/Users/gq2/git3/variantcalling/assets/vectorDB.tar.gz", + "default": "${projectDir}/assets/vectorDB.tar.gz", "description": "Path to directory or tar.gz archive for pre-built PacBio vector database.", "format": "file-path" }, From 45a7ba4d3b5a8856003cbf1cbb7a79725be41f17 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Thu, 7 Dec 2023 15:54:06 +0000 Subject: [PATCH 107/123] change maxRetries to 5. --- conf/base.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/base.config b/conf/base.config index c4ea647..e08d741 100644 --- a/conf/base.config +++ b/conf/base.config @@ -16,7 +16,7 @@ process { time = { check_max( 4.h * task.attempt, 'time' ) } errorStrategy = { task.exitStatus in ((130..145) + 104) ? 'retry' : 'finish' } - maxRetries = 1 + maxRetries = 5 maxErrors = '-1' // Process-specific resource requirements From 85215f56beff86cba592247b55d61c051ffa9bd6 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Sun, 17 Dec 2023 12:41:12 +0000 Subject: [PATCH 108/123] remove dev from the version --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index bb1f0f6..9cf2e35 100644 --- a/nextflow.config +++ b/nextflow.config @@ -215,7 +215,7 @@ manifest { description = """Variant calling pipeline for PacBio data using DeepVariant""" mainScript = 'main.nf' nextflowVersion = '!>=22.10.1' - version = '1.1.0-dev' + version = '1.1.0' doi = 'https://doi.org/10.5281/zenodo.7890527' } From 5f4419f57577b0657ffb2a3594ddaea6da46808c Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Mon, 18 Dec 2023 20:22:07 +0000 Subject: [PATCH 109/123] Add mermaid gitgraph workflow diagram --- docs/images/mermaid-diagram.png | Bin 0 -> 273704 bytes 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{'mainBranchName': 'BAM/CRAM'}} }%% + gitGraph TB: + commit id: "START" + branch Fasta order: 4 + commit id: "SAMTOOLS_FAIDX" + checkout BAM/CRAM + commit id: "SAMPLESHEET_CHECK" + branch AlignedReads order: 3 + branch UnAlignedReads order: 2 + commit id: "SAMTOOLS_COLLATE" + commit id: "SAMTOOLS_FASTA" + commit id: "BLAST_BLASTN" + commit id: "PACBIO_FILTER" + commit id: "PACBIO_SAMTOOLS_FILTER" + commit id: "SAMTOOLS_FASTQ" + commit id: "MINIMAP2_ALIGN" + commit id: "SAMTOOLS_MERGE_BY_SAMPLE 1" type: HIGHLIGHT tag: "output" + commit id: "SAMTOOLS_STATS" type: HIGHLIGHT tag: "output" + commit id: "SAMTOOLS_FLAGSTAT" type: HIGHLIGHT tag: "output" + commit id: "SAMTOOLS_IDXSTATS" type: HIGHLIGHT tag: "output" + checkout BAM/CRAM + merge UnAlignedReads + checkout Fasta + branch SplitFasta order: 5 + commit id: "FASTA_SPLIT" + branch DeepVariant order: 6 + checkout AlignedReads + merge Fasta + commit id: "SAMTOOLS_SORT" + commit id: "SAMTOOLS_MERGE_BY_SAMPLE 2" + checkout BAM/CRAM + merge AlignedReads + commit id: "SAMTOOLS_FILTER" + checkout DeepVariant + merge BAM/CRAM + commit id: "DEEPVARIANT" + commit id: "BCFTOOLS_CONCAT_VCF" type: HIGHLIGHT tag: "output" + branch "VCFtools" order: 7 + commit id: "VCFTOOLS_SITE_PI" type: HIGHLIGHT tag: "output" + commit id: "VCFTOOLS_HET" type: HIGHLIGHT tag: "output" + checkout DeepVariant + commit id: "BCFTOOLS_CONCAT_GVCF" type: HIGHLIGHT tag: "output" + checkout BAM/CRAM + merge VCFtools + merge DeepVariant + commit id: "DUMPSOFTWAREVERSIONS" type: HIGHLIGHT tag: "output" From 4522f72571291793f091f94542265da8121e2bf8 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 11:01:35 +0000 Subject: [PATCH 110/123] increase commit label size --- docs/images/mermaid-diagram.png | Bin 273704 -> 179251 bytes docs/images/mermaid-diagram.yaml | 27 +++++++++++++++++---------- 2 files changed, 17 insertions(+), 10 deletions(-) diff --git a/docs/images/mermaid-diagram.png b/docs/images/mermaid-diagram.png index 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commit id: "SAMTOOLS_MERGE_BY_SAMPLE 1" type: HIGHLIGHT tag: "output" - commit id: "SAMTOOLS_STATS" type: HIGHLIGHT tag: "output" - commit id: "SAMTOOLS_FLAGSTAT" type: HIGHLIGHT tag: "output" - commit id: "SAMTOOLS_IDXSTATS" type: HIGHLIGHT tag: "output" + commit id: "SAMTOOLS_MERGE_BY_SAMPLE 1" type: HIGHLIGHT + commit id: "SAMTOOLS_STATS" type: HIGHLIGHT + commit id: "SAMTOOLS_FLAGSTAT" type: HIGHLIGHT + commit id: "SAMTOOLS_IDXSTATS" type: HIGHLIGHT checkout BAM/CRAM merge UnAlignedReads checkout Fasta @@ -37,13 +44,13 @@ title: Sanger-tol/Variantcalling Workflow checkout DeepVariant merge BAM/CRAM commit id: "DEEPVARIANT" - commit id: "BCFTOOLS_CONCAT_VCF" type: HIGHLIGHT tag: "output" + commit id: "BCFTOOLS_CONCAT_VCF" type: HIGHLIGHT branch "VCFtools" order: 7 - commit id: "VCFTOOLS_SITE_PI" type: HIGHLIGHT tag: "output" - commit id: "VCFTOOLS_HET" type: HIGHLIGHT tag: "output" + commit id: "VCFTOOLS_SITE_PI" type: HIGHLIGHT + commit id: "VCFTOOLS_HET" type: HIGHLIGHT checkout DeepVariant - commit id: "BCFTOOLS_CONCAT_GVCF" type: HIGHLIGHT tag: "output" + commit id: "BCFTOOLS_CONCAT_GVCF" type: HIGHLIGHT checkout BAM/CRAM merge VCFtools merge DeepVariant - commit id: "DUMPSOFTWAREVERSIONS" type: HIGHLIGHT tag: "output" + commit id: "DUMPSOFTWAREVERSIONS" type: HIGHLIGHT From ec4e8eeba733621819a7b6aeca4da3b0fdd6e870 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 14:25:56 +0000 Subject: [PATCH 111/123] mermaid code mainly markdown --- docs/images/mermaid-diagram.md | 57 ++++++++++++++++++++++++++++++++ docs/images/mermaid-diagram.yaml | 56 ------------------------------- 2 files changed, 57 insertions(+), 56 deletions(-) create mode 100644 docs/images/mermaid-diagram.md delete mode 100644 docs/images/mermaid-diagram.yaml diff --git a/docs/images/mermaid-diagram.md b/docs/images/mermaid-diagram.md new file mode 100644 index 0000000..b447cae --- /dev/null +++ b/docs/images/mermaid-diagram.md @@ -0,0 +1,57 @@ +--- +title: Sanger-tol/Variantcalling Workflow +--- + +%%{ init: { +'gitGraph': {'mainBranchName': 'BAM/CRAM' +}, +'themeVariables': { +'commitLabelFontSize': '18px' +} +} +}%% +gitGraph TB: +commit id: "START" +branch Fasta order: 4 +commit id: "SAMTOOLS_FAIDX" +checkout BAM/CRAM +commit id: "SAMPLESHEET_CHECK" +branch AlignedReads order: 3 +branch UnAlignedReads order: 2 +commit id: "SAMTOOLS_COLLATE" +commit id: "SAMTOOLS_FASTA" +commit id: "BLAST_BLASTN" +commit id: "PACBIO_FILTER" +commit id: "PACBIO_SAMTOOLS_FILTER" +commit id: "SAMTOOLS_FASTQ" +commit id: "MINIMAP2_ALIGN" +commit id: "SAMTOOLS_MERGE_BY_SAMPLE 1" type: HIGHLIGHT +commit id: "SAMTOOLS_STATS" type: HIGHLIGHT +commit id: "SAMTOOLS_FLAGSTAT" type: HIGHLIGHT +commit id: "SAMTOOLS_IDXSTATS" type: HIGHLIGHT +checkout BAM/CRAM +merge UnAlignedReads +checkout Fasta +branch SplitFasta order: 5 +commit id: "FASTA_SPLIT" +branch DeepVariant order: 6 +checkout AlignedReads +merge Fasta +commit id: "SAMTOOLS_SORT" +commit id: "SAMTOOLS_MERGE_BY_SAMPLE 2" +checkout BAM/CRAM +merge AlignedReads +commit id: "SAMTOOLS_FILTER" +checkout DeepVariant +merge BAM/CRAM +commit id: "DEEPVARIANT" +commit id: "BCFTOOLS_CONCAT_VCF" type: HIGHLIGHT +branch "VCFtools" order: 7 +commit id: "VCFTOOLS_SITE_PI" type: HIGHLIGHT +commit id: "VCFTOOLS_HET" type: HIGHLIGHT +checkout DeepVariant +commit id: "BCFTOOLS_CONCAT_GVCF" type: HIGHLIGHT +checkout BAM/CRAM +merge VCFtools +merge DeepVariant +commit id: "DUMPSOFTWAREVERSIONS" type: HIGHLIGHT diff --git a/docs/images/mermaid-diagram.yaml b/docs/images/mermaid-diagram.yaml deleted file mode 100644 index e425064..0000000 --- a/docs/images/mermaid-diagram.yaml +++ /dev/null @@ -1,56 +0,0 @@ ---- -title: Sanger-tol/Variantcalling Workflow ---- -%%{ init: { - 'gitGraph': {'mainBranchName': 'BAM/CRAM' - }, - 'themeVariables': { - 'commitLabelFontSize': '18px' - } - } -}%% - gitGraph TB: - commit id: "START" - branch Fasta order: 4 - commit id: "SAMTOOLS_FAIDX" - checkout BAM/CRAM - commit id: "SAMPLESHEET_CHECK" - branch AlignedReads order: 3 - branch UnAlignedReads order: 2 - commit id: "SAMTOOLS_COLLATE" - commit id: "SAMTOOLS_FASTA" - commit id: "BLAST_BLASTN" - commit id: "PACBIO_FILTER" - commit id: "PACBIO_SAMTOOLS_FILTER" - commit id: "SAMTOOLS_FASTQ" - commit id: "MINIMAP2_ALIGN" - commit id: "SAMTOOLS_MERGE_BY_SAMPLE 1" type: HIGHLIGHT - commit id: "SAMTOOLS_STATS" type: HIGHLIGHT - commit id: "SAMTOOLS_FLAGSTAT" type: HIGHLIGHT - commit id: "SAMTOOLS_IDXSTATS" type: HIGHLIGHT - checkout BAM/CRAM - merge UnAlignedReads - checkout Fasta - branch SplitFasta order: 5 - commit id: "FASTA_SPLIT" - branch DeepVariant order: 6 - checkout AlignedReads - merge Fasta - commit id: "SAMTOOLS_SORT" - commit id: "SAMTOOLS_MERGE_BY_SAMPLE 2" - checkout BAM/CRAM - merge AlignedReads - commit id: "SAMTOOLS_FILTER" - checkout DeepVariant - merge BAM/CRAM - commit id: "DEEPVARIANT" - commit id: "BCFTOOLS_CONCAT_VCF" type: HIGHLIGHT - branch "VCFtools" order: 7 - commit id: "VCFTOOLS_SITE_PI" type: HIGHLIGHT - commit id: "VCFTOOLS_HET" type: HIGHLIGHT - checkout DeepVariant - commit id: "BCFTOOLS_CONCAT_GVCF" type: HIGHLIGHT - checkout BAM/CRAM - merge VCFtools - merge DeepVariant - commit id: "DUMPSOFTWAREVERSIONS" type: HIGHLIGHT From c1dd0e3384a3c1484bab7ecbbaff5971b2cbd5c1 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 14:26:05 +0000 Subject: [PATCH 112/123] change logs --- CHANGELOG.md | 46 ++++++++++++++++++++++++++++++++++++++++++++++ 1 file changed, 46 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index c2e0db5..62bb55c 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,6 +3,52 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). +## [[1.1.0](https://github.com/sanger-tol/variantcalling/releases/tag/1.1.0)] - Shang Tang - [2023-12-20] + +### Enhancements & fixes + +- Updated the CI procedure to use "sanger-tol" rather than "nf-core" names. +- Renamed Sanger related Github CI test workflows. +- nf-core template was updated from 2.7 to 2.8. +- Removed BAM/CRAM index files from the Sample sheets. +- Made fasta index file optional from the inputs. +- Imported PacBio readmapping sub-workflows from [sanger-tol/readmapping pipeline](https://github.com/sanger-tol/readmapping/). Therefore, the pipeline can run on unaligned BAM/CRAM samples now. +- Use VCFtools to calculate per site nucleotide diversity. +- Use VCFtools to calculate heterozygosity. + +### Parameters + +This release with the following initial parameters: + +| Old parameter | New parameter | +| ------------- | ------------------- | +| --gzi | | +| | --vector_db | +| | --align | +| | --include_positions | +| | --exclude_positions | + +> **NB:** Parameter has been **updated** if both old and new parameter information is present.
    **NB:** Parameter has been **added** if just the new parameter information is present.
    **NB:** Parameter has been **removed** if new parameter information isn't present. + +### Software dependencies + +Note, since the pipeline is using Nextflow DSL2, each process will be run with its own [Biocontainer](https://biocontainers.pro/#/registry). This means that on occasion it is entirely possible for the pipeline to be using different versions of the same tool. However, the overall software dependency changes compared to the last release have been listed below for reference. Only `Docker` or `Singularity` containers are supported, `conda` is not supported. + +| Dependency | Old version | New version | +| ----------- | ----------- | ----------- | +| DeepVariant | 1.4.0 | 1.5.0 | +| samtools | 1.16.1 | 1.17 | +| bcftools | 1.16.1 | 1.17 | +| python | 3.11.0 | 3.11.4 | +| vcftools | | 0.1.16 | +| blast | | 2.14.1+ | +| gunzip: | | 1.10 | +| minimap2 | | 2.24-r1122 | +| awk | | 5.1.0 | +| untar | | 1.30 | + +> **NB:** Dependency has been **updated** if both old and new version information is present.
    **NB:** Dependency has been **added** if just the new version information is present.
    **NB:** Dependency has been **removed** if version information isn't present. + ## [[1.0.0](https://github.com/sanger-tol/variantcalling/releases/tag/1.0.0)] - [2023-05-03] Initial release of sanger-tol/variantcalling, created with the [nf-core](https://nf-co.re/) tools. From ac88f507aa39709ea5af91211e1b2f574a4a11f8 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 14:44:28 +0000 Subject: [PATCH 113/123] update readme --- README.md | 13 +++++++++++-- 1 file changed, 11 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index ed207d1..3a344df 100644 --- a/README.md +++ b/README.md @@ -18,15 +18,19 @@ On release, automated continuous integration tests run the pipeline on a full-si ## Pipeline summary -The pipeline takes aligned PacBio sample reads (CRAM/BAM files) from a CSV file and the reference file in FASTA format, and then uses DeepVariant tool to make variant calling. +The pipeline takes aligned or unaligned PacBio sample reads (CRAM/BAM files) from a CSV file and the reference file in FASTA format, and then uses DeepVariant tool to make variant calling. Steps involved: - Split fasta file into smaller files, normally one sequence per file unless the sequences are too small. +- Align the reads if not aligned. - Merge input BAM/CRAM files together if they have the same sample names. - Filter out reads using the `-F 0x900` option to only retain the primary alignments. - Run DeepVariant using filtered BAM/CRAM files against each of split fasta files. - Merge all VCF and GVCF files generated by DeepVariant by sample together for each input BAM/CRAM file. +- Run VCFtools to calculate heterozygosity and per site nucleotide diversity. + + ## Quick Start @@ -37,7 +41,12 @@ Steps involved: 3. Download the pipeline and test it on a minimal dataset with a single command: ```bash + # for aligned reads nextflow run sanger-tol/variantcalling -profile test,YOURPROFILE --outdir + + # for unaligned reads + nextflow run sanger-tol/variantcalling -profile test_align,YOURPROFILE --align --outdir + ``` Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string. @@ -50,7 +59,7 @@ Steps involved: 4. Start running your own analysis! ```bash - nextflow run sanger-tol/variantcalling --input samplesheet.csv --outdir --fasta genome.fasta.gz --fai genome.fasta.gz.fai --gzi genome.fasta.gz.gzi -profile + nextflow run sanger-tol/variantcalling --input samplesheet.csv --outdir --fasta genome.fasta.gz -profile ``` ## Credits From 0be537c4b33fa42faf5afa1ed2032c9096031082 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 14:54:10 +0000 Subject: [PATCH 114/123] update citations --- CITATIONS.md | 16 ++++++++++++++++ 1 file changed, 16 insertions(+) diff --git a/CITATIONS.md b/CITATIONS.md index 923ff06..a492677 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -22,6 +22,22 @@ > Danecek, Petr, et al. “Twelve Years of SAMtools and BCFtools.” GigaScience, vol. 10, no. 2, Jan. 2021, https://doi.org/10.1093/gigascience/giab008. +- [Blast](https://pubmed.ncbi.nlm.nih.gov/20003500/) + + > Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K, Madden TL. BLAST+: architecture and applications. BMC Bioinformatics. 2009 Dec 15;10:421. doi: 10.1186/1471-2105-10-421. PMID: 20003500; PMCID: PMC2803857. + +- [BWA-MEM2](https://ieeexplore.ieee.org/document/8820962) + + > Vasimuddin Md, Misra S, Li H, Aluru S. Efficient Architecture-Aware Acceleration of BWA-MEM for Multicore Systems. 2019 IEEE International Parallel and Distributed Processing Symposium. 2019 May;314–24. doi: 10.1109/IPDPS.2019.00041. + +- [Minimap2](https://pubmed.ncbi.nlm.nih.gov/34623391/) + + > Li H. New strategies to improve minimap2 alignment accuracy. Bioinformatics. 2021 Oct 8;37(23):4572–4. doi: 10.1093/bioinformatics/btab705. Epub ahead of print. PMID: 34623391; PMCID: PMC8652018. + +- [VCFTools](https://pubmed.ncbi.nlm.nih.gov/21653522/) + + > Danecek P, Auton A, Abecasis G, et al.: The variant call format and VCFtools. Bioinformatics. 2011 Aug 1;27(15):2156-8. doi: 10.1093/bioinformatics/btr330. Epub 2011 Jun 7. PubMed PMID: 21653522; PubMed Central PMCID: PMC3137218. + ## Software packaging/containerisation tools - [Anaconda](https://anaconda.com) From 34605be31dd74200565cab9a5f22cafd65d0d8de Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 15:02:48 +0000 Subject: [PATCH 115/123] usage --- docs/usage.md | 6 ++++-- 1 file changed, 4 insertions(+), 2 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 46b74b3..131c5b8 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -2,7 +2,7 @@ ## Introduction -The pipeline takes aligned sample reads (CRAM/BAM files) from a CSV file and a reference file in FASTA format, and then use DeepVariant to call variants. +The pipeline takes aligned or unaliged sample reads (CRAM/BAM files) from a CSV file and a reference file in FASTA format, and then use DeepVariant to call variants. ## Samplesheet input @@ -47,7 +47,7 @@ An [example samplesheet](../assets/samplesheet.csv) has been provided with the p The typical command for running the pipeline is as follows: ```bash -nextflow run sanger-tol/variantcalling --input samplesheet.csv --outdir --fasta genome.fasta.gz --fai genome.fasta.gz.fai --gzi genome.fasta.gz.gzi -profile docker +nextflow run sanger-tol/variantcalling --input samplesheet.csv --outdir --fasta genome.fasta.gz -profile docker ``` This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles. @@ -63,6 +63,8 @@ work # Directory containing the nextflow working files The pipeline will split the input fasta file into smaller files to run DeepVariant parallel. You can set the minimum split fasta file size from the command line. For example to set the minimum size as 10K using `--split_fasta_cutoff 10000`. +If the input BAM/CRAM files are not aligned, please add `--align` in your command. + ### Updating the pipeline When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline: From b0b489a13cec18c98a9b61764197619013661dc2 Mon Sep 17 00:00:00 2001 From: Guoying Qi Date: Tue, 19 Dec 2023 16:04:21 +0000 Subject: [PATCH 116/123] output updated --- docs/output.md | 44 ++++++++++++++++++++++++++++++++++++++++++-- 1 file changed, 42 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index cfadaac..fa2258c 100644 --- a/docs/output.md +++ b/docs/output.md @@ -10,9 +10,49 @@ The directories listed below will be created in the results directory after the The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: +- [Readmapping Alignments](#readmapping-alignments) - Optional aligned CRAM files generated by minimap2 +- [Alignments Statistics](#alignments-statistics) Optional statistics files generated by samtools +- [VCFtools Processing](#vcftools-processing) Heterozygosity and per site nucleotide diversity calculated by VCFtools - [PacBio Variant Calling](#pacbio-variant-calling) - VCF and GVCF compressed files generated by DeepVariant - [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution +### Readmapping Alignments + +The unaligned PacBio read data is being filtered and aligned using `minimap2`. The CRAM files from the same sample will be merged. + +

    +Output files + +- `readmapping` + - Aligned CRAM files: `.pacbio..cram`. + - Aligned CRAM index files: `.pacbio..cram.crai`. + +
    + +### Alignments Statistics + +The statistics for the aligned CRAM files will be calculated using `samtools`. + +
    +Output files + +- `statistics` + - Comprehensive statistics from alignment file: `.pacbio..stats`. + - Number of alignments for each FLAG type: `.pacbio..flagstats`. + - Alignment summary statistics: `.pacbio..idxstats`. + +
    + +### VCFtools Processing + +
    +Output files + +- Heterozygosity generated by VCFtools: `.pacbio._deepvariant.vcf.het`. +- Per site nucleotide diversity calculated by VCFtools: `.pacbio._deepvariant.vcf.sites.pi`. + +
    + ### PacBio Variant Calling The aligned PacBio read data is used to call variants with DeepVariant. This is done by splitting the genome fasta file for speed efficiency. `BCFTOOLS` is used to combine the split `VCF` and `GVCF` files generated by `DEEPVARIANT`. @@ -21,8 +61,8 @@ The aligned PacBio read data is used to call variants with DeepVariant. This is Output files - `variant_calling` - - Compressed VCF files: `_deepvariant.vcf.gz`. - - Compressed GVCF files: `_deepvariant.g.vcf.gz`. + - Compressed VCF files: `.pacbio._deepvariant.vcf.gz`. + - Compressed GVCF files: `.pacbio._deepvariant.g.vcf.gz`.