From 79635f16b2cf35b1b40c88c5cb466f3ad1f2ce15 Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Tue, 5 Nov 2024 16:58:40 -0800 Subject: [PATCH 01/13] fixed formatting and added in asmstats to metaflye.wdl --- input.json | 4 +- jgi_assembly.wdl | 55 +++---- shortReads_assembly.wdl | 339 +++++++++++++++++++++------------------- 3 files changed, 206 insertions(+), 192 deletions(-) diff --git a/input.json b/input.json index 78bdb32..2b3ee69 100755 --- a/input.json +++ b/input.json @@ -1,7 +1,7 @@ { - "jgi_metaAssembly.input_files":["https://data.microbiomedata.org/data/test_data/11809.7.220839.TCCTGAG-ACTGCAT.fastq.gz"], + "jgi_metaAssembly.input_files":["/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz"], "jgi_metaAssembly.proj":"nmdc:503125_160870", "jgi_metaAssembly.memory": "105G", "jgi_metaAssembly.threads": "16", - "jgi_metaAssembly.shortRead": true + "jgi_metaAssembly.shortRead": false } diff --git a/jgi_assembly.wdl b/jgi_assembly.wdl index 54a3926..e6bef9c 100644 --- a/jgi_assembly.wdl +++ b/jgi_assembly.wdl @@ -1,10 +1,10 @@ version 1.0 import "shortReads_assembly.wdl" as srma import "make_interleaved_WDL/make_interleaved_reads.wdl" as int -# import "jgi_meta_wdl/metagenome_improved/metaflye.wdl" as lrma -import "https://code.jgi.doe.gov/BFoster/jgi_meta_wdl/-/raw/bc7c4371ea0fa83355bada341ec353b9feb3eff2/metagenome_improved/metaflye.wdl" as lrma +import "jgi_meta_wdl/metagenome_improved/metaflye.wdl" as lrma +# import "https://code.jgi.doe.gov/BFoster/jgi_meta_wdl/-/raw/bc7c4371ea0fa83355bada341ec353b9feb3eff2/metagenome_improved/metaflye.wdl" as lrma -workflow jgi_metaAssembly{ +workflow jgi_metaAssembly { input { Boolean shortRead String proj @@ -26,14 +26,14 @@ workflow jgi_metaAssembly{ if (shortRead) { - if (length(input_files) > 1){ - call int.make_interleaved_reads{ + if (length(input_files) > 1) { + call int.make_interleaved_reads { input: input_files = input_files, container = bbtools_container } } - call srma.jgi_metaASM{ + call srma.jgi_metaASM { input: memory = memory, threads = threads, @@ -45,7 +45,7 @@ workflow jgi_metaAssembly{ } if (!shortRead) { - call lrma.metaflye{ + call lrma.metaflye { input: proj = proj, input_fastq = input_files, @@ -58,7 +58,7 @@ workflow jgi_metaAssembly{ samtools_container = samtools_container, bbtools_container = bbtools_container } - call finish_lrasm{ + call finish_lrasm { input: proj = proj, prefix = prefix, @@ -98,33 +98,34 @@ workflow jgi_metaAssembly{ File? sr_bam=jgi_metaASM.bam File? sr_samgz=jgi_metaASM.samgz File? sr_covstats=jgi_metaASM.covstats - File? sr_asmstats=jgi_metaASM.asmstats File? sr_asminfo=jgi_metaASM.asminfo File? sr_bbcms_fq = jgi_metaASM.bbcms_fastq - + + #Both + File? stats = if (shortRead) then jgi_metaASM.asmstats else metaflye.asmstats } } task finish_lrasm { input { - File contigs - File bam - File scaffolds - File agp - File legend - File basecov - File sam - File output_file - File stats - File summary_stats - File pileup_out - String container - String proj - String prefix - String orig_prefix="scaffold" - String sed="s/~{orig_prefix}_/~{proj}_/g" - # String start + File contigs + File bam + File scaffolds + File agp + File legend + File basecov + File sam + File output_file + File stats + File summary_stats + File pileup_out + String container + String proj + String prefix + String orig_prefix="scaffold" + String sed="s/~{orig_prefix}_/~{proj}_/g" + # String start } command<<< diff --git a/shortReads_assembly.wdl b/shortReads_assembly.wdl index 5452326..8ce8845 100755 --- a/shortReads_assembly.wdl +++ b/shortReads_assembly.wdl @@ -1,6 +1,6 @@ version 1.0 workflow jgi_metaASM { - input{ + input { # String? outdir String? memory String? threads @@ -13,7 +13,7 @@ workflow jgi_metaASM { String spades_container="microbiomedata/spades:3.15.0" String worflowmeta_container="microbiomedata/workflowmeta:1.1.1" Boolean paired = true - } + } call stage { input: @@ -117,49 +117,51 @@ workflow jgi_metaASM { } task stage { - input{ - String container - String? input_file - String memory = "4G" - String target = "staged.fastq.gz" - String output1 = "input.left.fastq.gz" - String output2 = "input.right.fastq.gz"} - - command <<< - set -euo pipefail - if [ $( echo ~{input_file}|egrep -c "https*:") -gt 0 ] ; then - wget ~{input_file} -O ~{target} - else - ln -s ~{input_file} ~{target} || cp ~{input_file} ~{target} - fi + input { + String container + String? input_file + String memory = "4G" + String target = "staged.fastq.gz" + String output1 = "input.left.fastq.gz" + String output2 = "input.right.fastq.gz" + } + + command<<< + + set -euo pipefail + if [ $( echo ~{input_file}|egrep -c "https*:") -gt 0 ] ; then + wget ~{input_file} -O ~{target} + else + ln -s ~{input_file} ~{target} || cp ~{input_file} ~{target} + fi reformat.sh \ ~{if (defined(memory)) then "-Xmx" + memory else "-Xmx10G" }\ in=~{target} \ out1=~{output1} \ out2=~{output2} - # Capture the start time - date --iso-8601=seconds > start.txt - - >>> - - output{ - Array[File] assembly_input = [output1, output2] - String start = read_string("start.txt") - } - runtime { - cpu: 2 - maxRetries: 1 - docker: container - } + # Capture the start time + date --iso-8601=seconds > start.txt + + >>> + + output { + Array[File] assembly_input = [output1, output2] + String start = read_string("start.txt") + } + runtime { + cpu: 2 + maxRetries: 1 + docker: container + } } task make_info_file { - input{ - File assy_info - File bbcms_info - String prefix - String container + input { + File assy_info + File bbcms_info + String prefix + String container } command<<< @@ -178,6 +180,7 @@ task make_info_file { output { File asminfo = "~{prefix}_metaAsm.info" } + runtime { memory: "1 GiB" cpu: 1 @@ -187,22 +190,23 @@ task make_info_file { } task finish_asm { - input{ - File fasta - File scaffold - File? agp - File bam - File? samgz - File? covstats - File asmstats - File bbcms_fastq - String container - String proj - String prefix - String orig_prefix="scaffold" - String sed="s/~{orig_prefix}_/~{proj}_/g" - # String start + input { + File fasta + File scaffold + File? agp + File bam + File? samgz + File? covstats + File asmstats + File bbcms_fastq + String container + String proj + String prefix + String orig_prefix="scaffold" + String sed="s/~{orig_prefix}_/~{proj}_/g" + # String start } + command<<< set -euo pipefail @@ -249,32 +253,28 @@ task finish_asm { } -task read_mapping_pairs{ - input{ - Array[File] reads - File ref - String container - String? memory - String? threads - Boolean paired = true - String bbmap_interleaved_flag = if paired then 'interleaved=true' else 'interleaved=false' - - String filename_unsorted="pairedMapped.bam" - String filename_outsam="pairedMapped.sam.gz" - String filename_sorted="pairedMapped_sorted.bam" - String filename_sorted_idx="pairedMapped_sorted.bam.bai" - String filename_bamscript="to_bam.sh" - String filename_cov="covstats.txt" - String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" - String jvm_threads=select_first([threads,system_cpu]) +task read_mapping_pairs { + input { + Array[File] reads + File ref + String container + String? memory + String? threads + Boolean paired = true + String bbmap_interleaved_flag = if paired then 'interleaved=true' else 'interleaved=false' + + String filename_unsorted="pairedMapped.bam" + String filename_outsam="pairedMapped.sam.gz" + String filename_sorted="pairedMapped_sorted.bam" + String filename_sorted_idx="pairedMapped_sorted.bam.bai" + String filename_bamscript="to_bam.sh" + String filename_cov="covstats.txt" + String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" + String jvm_threads=select_first([threads,system_cpu]) } - runtime { - docker: container - memory: "120 GiB" - cpu: 16 - maxRetries: 1 - } - command{ + + command<<< + set -euo pipefail if [[ ~{reads[0]} == *.gz ]] ; then cat ~{sep=" " reads} > infile.fastq.gz @@ -314,33 +314,36 @@ task read_mapping_pairs{ ln -s ~{filename_cov} mapping_stats.txt rm $mapping_input - } - output{ - File outbamfile = filename_sorted - File outbamfileidx = filename_sorted_idx - File outcovfile = filename_cov - File outsamfile = filename_outsam - } -} -task create_agp { - input{ - File scaffolds_in - String? memory - String container - String rename_contig_prefix - String prefix="assembly" - String filename_contigs="~{prefix}_contigs.fna" - String filename_scaffolds="~{prefix}_scaffolds.fna" - String filename_agp="~{prefix}.agp" - String filename_legend="~{prefix}_scaffolds.legend" + >>> + output { + File outbamfile = filename_sorted + File outbamfileidx = filename_sorted_idx + File outcovfile = filename_cov + File outsamfile = filename_outsam } runtime { - docker: container - memory: "120 GiB" + docker: container + memory: "120 GiB" cpu: 16 - } - command{ + maxRetries: 1 + } +} + +task create_agp { + input { + File scaffolds_in + String? memory + String container + String rename_contig_prefix + String prefix="assembly" + String filename_contigs="~{prefix}_contigs.fna" + String filename_scaffolds="~{prefix}_scaffolds.fna" + String filename_agp="~{prefix}.agp" + String filename_legend="~{prefix}_scaffolds.legend" + } + + command<<< set -euo pipefail fungalrelease.sh \ ~{if (defined(memory)) then "-Xmx" + memory else "-Xmx105G" } \ @@ -361,35 +364,39 @@ task create_agp { fi bbstats.sh format=8 in=~{filename_scaffolds} out=stats.json sed -i 's/l_gt50k/l_gt50K/g' stats.json + + >>> + + output { + File outcontigs = filename_contigs + File outscaffolds = filename_scaffolds + File outagp = filename_agp + File outstats = "stats.json" + File outlegend = filename_legend } - output{ - File outcontigs = filename_contigs - File outscaffolds = filename_scaffolds - File outagp = filename_agp - File outstats = "stats.json" - File outlegend = filename_legend + + runtime { + docker: container + memory: "120 GiB" + cpu: 16 } } task assy { - input{ - File infile1 - File infile2 - String container - String? threads - String outprefix="spades3" - String filename_outfile="~{outprefix}/scaffolds.fasta" - String filename_spadeslog ="~{outprefix}/spades.log" - String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" - String spades_cpu=select_first([threads,system_cpu]) - Boolean paired = true + input { + File infile1 + File infile2 + String container + String? threads + String outprefix="spades3" + String filename_outfile="~{outprefix}/scaffolds.fasta" + String filename_spadeslog ="~{outprefix}/spades.log" + String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" + String spades_cpu=select_first([threads,system_cpu]) + Boolean paired = true } - runtime { - docker: container - memory: "120 GiB" - cpu: 16 - } - command{ + + command <<< set -euo pipefail if ~{paired}; then spades.py \ @@ -410,44 +417,45 @@ task assy { -t ~{spades_cpu} \ -s ~{infile1} fi - } - output { - File out = filename_outfile - File outlog = filename_spadeslog - } + >>> + + output { + File out = filename_outfile + File outlog = filename_spadeslog + } + runtime { + docker: container + memory: "120 GiB" + cpu: 16 + } } task bbcms { - input{ - Array[File] input_files - String container - String? memory - Boolean paired = true - String filename_outfile="input.corr.fastq.gz" - String filename_outfile1="input.corr.left.fastq.gz" - String filename_outfile2="input.corr.right.fastq.gz" - String filename_readlen="readlen.txt" - String filename_outlog="stdout.log" - String filename_errlog="stderr.log" - String filename_kmerfile="unique31mer.txt" - String filename_counts="counts.metadata.json" + input { + Array[File] input_files + String container + String? memory + Boolean paired = true + String filename_outfile="input.corr.fastq.gz" + String filename_outfile1="input.corr.left.fastq.gz" + String filename_outfile2="input.corr.right.fastq.gz" + String filename_readlen="readlen.txt" + String filename_outlog="stdout.log" + String filename_errlog="stderr.log" + String filename_kmerfile="unique31mer.txt" + String filename_counts="counts.metadata.json" } - runtime { - docker: container - memory: "120 GiB" - cpu: 16 - } - command { + command<<< set -euo pipefail if file --mime -b ~{input_files[0]} | grep gzip; then - cat ~{sep=" " input_files} > infile.fastq.gz - export bbcms_input="infile.fastq.gz" + cat ~{sep=" " input_files} > infile.fastq.gz + export bbcms_input="infile.fastq.gz" fi if file --mime -b ~{input_files[0]} | grep plain; then - cat ~{sep=" " input_files} > infile.fastq - export bbcms_input="infile.fastq" + cat ~{sep=" " input_files} > infile.fastq + export bbcms_input="infile.fastq" fi bbcms.sh \ @@ -477,19 +485,24 @@ task bbcms { out=~{filename_readlen} rm $bbcms_input - - } - output { - File out = filename_outfile - File out1 = if paired then filename_outfile1 else filename_outfile - File out2 = if paired then filename_outfile2 else filename_outfile - File outreadlen = filename_readlen - File stdout = filename_outlog - File stderr = filename_errlog - File outcounts = filename_counts - File outkmer = filename_kmerfile - - } + + >>> + + output { + File out = filename_outfile + File out1 = if paired then filename_outfile1 else filename_outfile + File out2 = if paired then filename_outfile2 else filename_outfile + File outreadlen = filename_readlen + File stdout = filename_outlog + File stderr = filename_errlog + File outcounts = filename_counts + File outkmer = filename_kmerfile + } + runtime { + docker: container + memory: "120 GiB" + cpu: 16 + } } # task make_output{ From 64f846727413f633d4adb2d3713db53b436deb4e Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Thu, 7 Nov 2024 11:11:57 -0800 Subject: [PATCH 02/13] added stats.json as an output for task finish_lrasm --- jgi_assembly.wdl | 24 +++++++++++++++--------- 1 file changed, 15 insertions(+), 9 deletions(-) diff --git a/jgi_assembly.wdl b/jgi_assembly.wdl index e6bef9c..6cf2bd0 100644 --- a/jgi_assembly.wdl +++ b/jgi_assembly.wdl @@ -1,8 +1,8 @@ version 1.0 import "shortReads_assembly.wdl" as srma import "make_interleaved_WDL/make_interleaved_reads.wdl" as int -import "jgi_meta_wdl/metagenome_improved/metaflye.wdl" as lrma -# import "https://code.jgi.doe.gov/BFoster/jgi_meta_wdl/-/raw/bc7c4371ea0fa83355bada341ec353b9feb3eff2/metagenome_improved/metaflye.wdl" as lrma +# import "metaAssembly/metagenome_improved/metaflye.wdl" as lrma +import "https://code.jgi.doe.gov/BFoster/jgi_meta_wdl/-/raw/bc7c4371ea0fa83355bada341ec353b9feb3eff2/metagenome_improved/metaflye.wdl" as lrma workflow jgi_metaAssembly { input { @@ -102,7 +102,7 @@ workflow jgi_metaAssembly { File? sr_bbcms_fq = jgi_metaASM.bbcms_fastq #Both - File? stats = if (shortRead) then jgi_metaASM.asmstats else metaflye.asmstats + File? stats = if (shortRead) then jgi_metaASM.asmstats else finish_lrasm.asm_stats } } @@ -143,12 +143,17 @@ task finish_lrasm { cat ~{basecov} | sed ~{sed} > ~{prefix}_contigs.sorted.bam.pileup.basecov cat ~{pileup_out} | sed ~{sed} > ~{prefix}_contigs.sorted.bam.pileup.out - ## Bam file - samtools view -h ~{bam} | sed ~{sed} | \ - samtools view -hb -o ~{prefix}_pairedMapped_sorted.bam - ## Sam.gz file - samtools view -h ~{sam} | sed ~{sed} | \ - gzip -c - > ~{prefix}_pairedMapped.sam.gz + ## Bam file + samtools view -h ~{bam} | sed ~{sed} | \ + samtools view -hb -o ~{prefix}_pairedMapped_sorted.bam + ## Sam.gz file + samtools view -h ~{sam} | sed ~{sed} | \ + gzip -c - > ~{prefix}_pairedMapped.sam.gz + + # stats file + bbstats.sh format=8 in=~{scaffolds} out=stats.json + sed -i 's/l_gt50k/l_gt50K/g' stats.json + cat stats.json |jq 'del(.filename)' > stats.json >>> output { @@ -163,6 +168,7 @@ task finish_lrasm { File final_stats = "~{prefix}_contigs.sam.stats" File final_summary_stats = "~{prefix}_summary.stats" File final_pileup_out = "~{prefix}_contigs.sorted.bam.pileup.out" + File asm_stats = "stats.json" } runtime { From 7766db87bff8d6493d671ddf081bf8039561eba7 Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Thu, 7 Nov 2024 12:24:27 -0800 Subject: [PATCH 03/13] worflowmeta_contiainer -> workflowmeta_container --- shortReads_assembly.wdl | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/shortReads_assembly.wdl b/shortReads_assembly.wdl index 8ce8845..7609043 100755 --- a/shortReads_assembly.wdl +++ b/shortReads_assembly.wdl @@ -11,7 +11,7 @@ workflow jgi_metaASM { # Float uniquekmer=1000 String bbtools_container="microbiomedata/bbtools:38.96" String spades_container="microbiomedata/spades:3.15.0" - String worflowmeta_container="microbiomedata/workflowmeta:1.1.1" + String workflowmeta_container="microbiomedata/workflowmeta:1.1.1" Boolean paired = true } @@ -65,7 +65,7 @@ workflow jgi_metaASM { proj=proj, prefix=prefix, # start=stage.start, - container=worflowmeta_container, + container=workflowmeta_container, fasta=create_agp.outcontigs, scaffold=create_agp.outscaffolds, agp=create_agp.outagp, @@ -93,7 +93,7 @@ workflow jgi_metaASM { # samgz_name=basename(finish_asm.outcontigs), # covstats_name=basename(finish_asm.outcontigs), # asmstats_name=basename(finish_asm.outcontigs), - # container = worflowmeta_container + # container = workflowmeta_container # } output { From d8b10a9aa9db6f799d7f7e51be3e20fd7ed86d9c Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Thu, 7 Nov 2024 14:05:48 -0800 Subject: [PATCH 04/13] Updated bbtools container --- jgi_assembly.wdl | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/jgi_assembly.wdl b/jgi_assembly.wdl index 6cf2bd0..34e0bb2 100644 --- a/jgi_assembly.wdl +++ b/jgi_assembly.wdl @@ -21,7 +21,7 @@ workflow jgi_metaAssembly { String minimap2_container = "staphb/minimap2:2.25" String minimap2_parameters = "-a -x map-hifi -t 32" String samtools_container = "staphb/samtools:1.18" - String bbtools_container = "microbiomedata/bbtools:38.96" + String bbtools_container = "microbiomedata/bbtools:39.03" } From f8dddd64b5017bf0cb4ad0e3cade43c46ced1ab5 Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Fri, 8 Nov 2024 07:59:31 -0800 Subject: [PATCH 05/13] reverted bbtools container for shortreads --- jgi_assembly.wdl | 4 +- shortReads_assembly.wdl | 315 ++++++++++++++++++++-------------------- 2 files changed, 156 insertions(+), 163 deletions(-) diff --git a/jgi_assembly.wdl b/jgi_assembly.wdl index 34e0bb2..4c6a913 100644 --- a/jgi_assembly.wdl +++ b/jgi_assembly.wdl @@ -30,7 +30,7 @@ workflow jgi_metaAssembly { call int.make_interleaved_reads { input: input_files = input_files, - container = bbtools_container + container = "microbiomedata/bbtools:38.96" } } call srma.jgi_metaASM { @@ -39,7 +39,7 @@ workflow jgi_metaAssembly { threads = threads, input_file = if length(input_files) > 1 then make_interleaved_reads.interleaved_fastq else input_files[0], proj = proj, - bbtools_container = bbtools_container + bbtools_container = "microbiomedata/bbtools:38.96" } diff --git a/shortReads_assembly.wdl b/shortReads_assembly.wdl index 7609043..511b845 100755 --- a/shortReads_assembly.wdl +++ b/shortReads_assembly.wdl @@ -10,10 +10,10 @@ workflow jgi_metaASM { String rename_contig_prefix="scaffold" # Float uniquekmer=1000 String bbtools_container="microbiomedata/bbtools:38.96" - String spades_container="microbiomedata/spades:3.15.0" + String spades_container="staphb/spades:4.0.0" String workflowmeta_container="microbiomedata/workflowmeta:1.1.1" Boolean paired = true - } + } call stage { input: @@ -117,51 +117,50 @@ workflow jgi_metaASM { } task stage { - input { - String container - String? input_file - String memory = "4G" - String target = "staged.fastq.gz" - String output1 = "input.left.fastq.gz" - String output2 = "input.right.fastq.gz" - } - - command<<< - - set -euo pipefail - if [ $( echo ~{input_file}|egrep -c "https*:") -gt 0 ] ; then - wget ~{input_file} -O ~{target} - else - ln -s ~{input_file} ~{target} || cp ~{input_file} ~{target} - fi + input { + String container + String? input_file + String memory = "4G" + String target = "staged.fastq.gz" + String output1 = "input.left.fastq.gz" + String output2 = "input.right.fastq.gz" + } + + command <<< + set -euo pipefail + if [ $( echo ~{input_file}|egrep -c "https*:") -gt 0 ] ; then + wget ~{input_file} -O ~{target} + else + ln -s ~{input_file} ~{target} || cp ~{input_file} ~{target} + fi reformat.sh \ ~{if (defined(memory)) then "-Xmx" + memory else "-Xmx10G" }\ in=~{target} \ out1=~{output1} \ out2=~{output2} - # Capture the start time - date --iso-8601=seconds > start.txt - - >>> - - output { - Array[File] assembly_input = [output1, output2] - String start = read_string("start.txt") - } - runtime { - cpu: 2 - maxRetries: 1 - docker: container - } + # Capture the start time + date --iso-8601=seconds > start.txt + + >>> + + output { + Array[File] assembly_input = [output1, output2] + String start = read_string("start.txt") + } + runtime { + cpu: 2 + maxRetries: 1 + docker: container + } } task make_info_file { - input { - File assy_info - File bbcms_info - String prefix - String container + input{ + File assy_info + File bbcms_info + String prefix + String container } command<<< @@ -180,7 +179,6 @@ task make_info_file { output { File asminfo = "~{prefix}_metaAsm.info" } - runtime { memory: "1 GiB" cpu: 1 @@ -191,20 +189,20 @@ task make_info_file { task finish_asm { input { - File fasta - File scaffold - File? agp - File bam - File? samgz - File? covstats - File asmstats - File bbcms_fastq - String container - String proj - String prefix - String orig_prefix="scaffold" - String sed="s/~{orig_prefix}_/~{proj}_/g" - # String start + File fasta + File scaffold + File? agp + File bam + File? samgz + File? covstats + File asmstats + File bbcms_fastq + String container + String proj + String prefix + String orig_prefix="scaffold" + String sed="s/~{orig_prefix}_/~{proj}_/g" + # String start } command<<< @@ -253,28 +251,32 @@ task finish_asm { } -task read_mapping_pairs { +task read_mapping_pairs{ input { - Array[File] reads - File ref - String container - String? memory - String? threads - Boolean paired = true - String bbmap_interleaved_flag = if paired then 'interleaved=true' else 'interleaved=false' - - String filename_unsorted="pairedMapped.bam" - String filename_outsam="pairedMapped.sam.gz" - String filename_sorted="pairedMapped_sorted.bam" - String filename_sorted_idx="pairedMapped_sorted.bam.bai" - String filename_bamscript="to_bam.sh" - String filename_cov="covstats.txt" - String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" - String jvm_threads=select_first([threads,system_cpu]) + Array[File] reads + File ref + String container + String? memory + String? threads + Boolean paired = true + String bbmap_interleaved_flag = if paired then 'interleaved=true' else 'interleaved=false' + + String filename_unsorted="pairedMapped.bam" + String filename_outsam="pairedMapped.sam.gz" + String filename_sorted="pairedMapped_sorted.bam" + String filename_sorted_idx="pairedMapped_sorted.bam.bai" + String filename_bamscript="to_bam.sh" + String filename_cov="covstats.txt" + String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" + String jvm_threads=select_first([threads,system_cpu]) } - + runtime { + docker: container + memory: "120 GiB" + cpu: 16 + maxRetries: 1 + } command<<< - set -euo pipefail if [[ ~{reads[0]} == *.gz ]] ; then cat ~{sep=" " reads} > infile.fastq.gz @@ -317,32 +319,30 @@ task read_mapping_pairs { >>> output { - File outbamfile = filename_sorted - File outbamfileidx = filename_sorted_idx - File outcovfile = filename_cov - File outsamfile = filename_outsam - } - runtime { - docker: container - memory: "120 GiB" - cpu: 16 - maxRetries: 1 - } + File outbamfile = filename_sorted + File outbamfileidx = filename_sorted_idx + File outcovfile = filename_cov + File outsamfile = filename_outsam + } } task create_agp { input { - File scaffolds_in - String? memory - String container - String rename_contig_prefix - String prefix="assembly" - String filename_contigs="~{prefix}_contigs.fna" - String filename_scaffolds="~{prefix}_scaffolds.fna" - String filename_agp="~{prefix}.agp" - String filename_legend="~{prefix}_scaffolds.legend" + File scaffolds_in + String? memory + String container + String rename_contig_prefix + String prefix="assembly" + String filename_contigs="~{prefix}_contigs.fna" + String filename_scaffolds="~{prefix}_scaffolds.fna" + String filename_agp="~{prefix}.agp" + String filename_legend="~{prefix}_scaffolds.legend" } - + runtime { + docker: container + memory: "120 GiB" + cpu: 16 + } command<<< set -euo pipefail fungalrelease.sh \ @@ -368,35 +368,33 @@ task create_agp { >>> output { - File outcontigs = filename_contigs - File outscaffolds = filename_scaffolds - File outagp = filename_agp - File outstats = "stats.json" - File outlegend = filename_legend - } - - runtime { - docker: container - memory: "120 GiB" - cpu: 16 + File outcontigs = filename_contigs + File outscaffolds = filename_scaffolds + File outagp = filename_agp + File outstats = "stats.json" + File outlegend = filename_legend } } task assy { - input { - File infile1 - File infile2 - String container - String? threads - String outprefix="spades3" - String filename_outfile="~{outprefix}/scaffolds.fasta" - String filename_spadeslog ="~{outprefix}/spades.log" - String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" - String spades_cpu=select_first([threads,system_cpu]) - Boolean paired = true + input{ + File infile1 + File infile2 + String container + String? threads + String outprefix="spades3" + String filename_outfile="~{outprefix}/scaffolds.fasta" + String filename_spadeslog ="~{outprefix}/spades.log" + String system_cpu="$(grep \"model name\" /proc/cpuinfo | wc -l)" + String spades_cpu=select_first([threads,system_cpu]) + Boolean paired = true } - - command <<< + runtime { + docker: container + memory: "120 GiB" + cpu: 16 + } + command <<< set -euo pipefail if ~{paired}; then spades.py \ @@ -419,43 +417,43 @@ task assy { fi >>> - output { - File out = filename_outfile - File outlog = filename_spadeslog - } - runtime { - docker: container - memory: "120 GiB" - cpu: 16 - } + output { + File out = filename_outfile + File outlog = filename_spadeslog + } } task bbcms { - input { - Array[File] input_files - String container - String? memory - Boolean paired = true - String filename_outfile="input.corr.fastq.gz" - String filename_outfile1="input.corr.left.fastq.gz" - String filename_outfile2="input.corr.right.fastq.gz" - String filename_readlen="readlen.txt" - String filename_outlog="stdout.log" - String filename_errlog="stderr.log" - String filename_kmerfile="unique31mer.txt" - String filename_counts="counts.metadata.json" + input{ + Array[File] input_files + String container + String? memory + Boolean paired = true + String filename_outfile="input.corr.fastq.gz" + String filename_outfile1="input.corr.left.fastq.gz" + String filename_outfile2="input.corr.right.fastq.gz" + String filename_readlen="readlen.txt" + String filename_outlog="stdout.log" + String filename_errlog="stderr.log" + String filename_kmerfile="unique31mer.txt" + String filename_counts="counts.metadata.json" } + runtime { + docker: container + memory: "120 GiB" + cpu: 16 + } command<<< set -euo pipefail if file --mime -b ~{input_files[0]} | grep gzip; then - cat ~{sep=" " input_files} > infile.fastq.gz - export bbcms_input="infile.fastq.gz" + cat ~{sep=" " input_files} > infile.fastq.gz + export bbcms_input="infile.fastq.gz" fi if file --mime -b ~{input_files[0]} | grep plain; then - cat ~{sep=" " input_files} > infile.fastq - export bbcms_input="infile.fastq" + cat ~{sep=" " input_files} > infile.fastq + export bbcms_input="infile.fastq" fi bbcms.sh \ @@ -485,24 +483,20 @@ task bbcms { out=~{filename_readlen} rm $bbcms_input - + >>> - output { - File out = filename_outfile - File out1 = if paired then filename_outfile1 else filename_outfile - File out2 = if paired then filename_outfile2 else filename_outfile - File outreadlen = filename_readlen - File stdout = filename_outlog - File stderr = filename_errlog - File outcounts = filename_counts - File outkmer = filename_kmerfile - } - runtime { - docker: container - memory: "120 GiB" - cpu: 16 - } + output { + File out = filename_outfile + File out1 = if paired then filename_outfile1 else filename_outfile + File out2 = if paired then filename_outfile2 else filename_outfile + File outreadlen = filename_readlen + File stdout = filename_outlog + File stderr = filename_errlog + File outcounts = filename_counts + File outkmer = filename_kmerfile + + } } # task make_output{ @@ -546,5 +540,4 @@ task bbcms { # File? outcovstats = "~{outdir}/~{covstats_name}" # File? outasmstats = "~{outdir}/~{asmstats_name}" # } -# } - +# } \ No newline at end of file From 061e646156b8dec2cc9bdfd759c33773ad9351e6 Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Fri, 8 Nov 2024 08:25:05 -0800 Subject: [PATCH 06/13] formatting issues --- jgi_assembly.wdl | 67 ++++++++++++++++++++++++------------------------ 1 file changed, 33 insertions(+), 34 deletions(-) diff --git a/jgi_assembly.wdl b/jgi_assembly.wdl index 7d64a39..a18c20b 100644 --- a/jgi_assembly.wdl +++ b/jgi_assembly.wdl @@ -28,53 +28,52 @@ workflow jgi_metaAssembly { if (shortRead) { if (length(input_files) > 1) { call int.make_interleaved_reads { - input: - input_files = input_files, - container = "microbiomedata/bbtools:38.96" + input: + input_files = input_files, + container = "microbiomedata/bbtools:38.96" } } call srma.jgi_metaASM { input: - memory = memory, - threads = threads, - input_file = if length(input_files) > 1 then make_interleaved_reads.interleaved_fastq else input_files[0], - proj = proj, - bbtools_container = "microbiomedata/bbtools:38.96" - spades_container = spades_container - + memory = memory, + threads = threads, + input_file = if length(input_files) > 1 then make_interleaved_reads.interleaved_fastq else input_files[0], + proj = proj, + bbtools_container = "microbiomedata/bbtools:38.96", + spades_container = spades_container } } if (!shortRead) { call lrma.metaflye { input: - proj = proj, - input_fastq = input_files, - flye_container = flye_container, - flye_parameters = flye_parameters, - smrtlink_container = smrtlink_container, - racon_container = racon_container, - minimap2_container = minimap2_container, - minimap2_parameters = minimap2_parameters, - samtools_container = samtools_container, - bbtools_container = bbtools_container + proj = proj, + input_fastq = input_files, + flye_container = flye_container, + flye_parameters = flye_parameters, + smrtlink_container = smrtlink_container, + racon_container = racon_container, + minimap2_container = minimap2_container, + minimap2_parameters = minimap2_parameters, + samtools_container = samtools_container, + bbtools_container = bbtools_container } call finish_lrasm { input: - proj = proj, - prefix = prefix, - container = bbtools_container, - contigs = metaflye.final_contigs, - bam = metaflye.final_bam, - scaffolds = metaflye.final_scaffolds, - agp = metaflye.final_agp, - legend = metaflye.final_legend, - basecov = metaflye.final_basecov, - sam = metaflye.final_sam, - output_file = metaflye.final_output_file, - stats = metaflye.final_stats, - summary_stats = metaflye.final_summary_stats, - pileup_out = metaflye.final_pileup_out + proj = proj, + prefix = prefix, + container = bbtools_container, + contigs = metaflye.final_contigs, + bam = metaflye.final_bam, + scaffolds = metaflye.final_scaffolds, + agp = metaflye.final_agp, + legend = metaflye.final_legend, + basecov = metaflye.final_basecov, + sam = metaflye.final_sam, + output_file = metaflye.final_output_file, + stats = metaflye.final_stats, + summary_stats = metaflye.final_summary_stats, + pileup_out = metaflye.final_pileup_out } } output { From 859c0619458b94fb060ebfcfa537757c990c6fee Mon Sep 17 00:00:00 2001 From: Valerie Li <147084799+vlilanl@users.noreply.github.com> Date: Tue, 12 Nov 2024 08:57:16 -0700 Subject: [PATCH 07/13] Update index.rst --- docs/index.rst | 87 +++++++++++++++++++++++++++++++++----------------- 1 file changed, 57 insertions(+), 30 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index be63c31..ba0becf 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -1,4 +1,4 @@ -Metagenome Assembly Workflow (v1.0.2) +Metagenome Assembly Workflow (v1.0.6) ======================================== .. image:: workflow_assembly.png @@ -8,7 +8,7 @@ Metagenome Assembly Workflow (v1.0.2) Workflow Overview ----------------- -This workflow takes in paired-end Illumina reads in interleaved format and performs error correction, then reformats the interleaved file into two FASTQ files for downstream tasks using bbcms (BBTools). The corrected reads are assembled using metaSPAdes. After assembly, the reads are mapped back to contigs by bbmap (BBTools) for coverage information. The .wdl (Workflow Description Language) file includes five tasks, *bbcms*, *assy*, *create_agp*, *read_mapping_pairs*, and *make_output*. +This workflow takes in paired-end Illumina short reads or paired-end PacBio long reads in interleaved format and performs error correction, then reformats the interleaved file into two FASTQ files for downstream tasks using bbcms (BBTools). The corrected reads are assembled using metaSPAdes. After assembly, the reads are mapped back to contigs by bbmap (BBTools) for coverage information. The .wdl (Workflow Description Language) file includes five tasks, *bbcms*, *assy*, *create_agp*, *read_mapping_pairs*, and *make_output*. 1. The *bbcms* task takes in interleaved FASTQ inputs and performs error correction and reformats the interleaved fastq into two output FASTQ files for paired-end reads for the next tasks. 2. The *assy* task performs metaSPAdes assembly @@ -62,6 +62,8 @@ Sample dataset(s) - large dataset: `Zymobiomics mock-community DNA control (22G) `_ . You can find input/output in the downloaded tar gz file. +- long reads dataset: `/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz` + Zymobiomics mock-community DNA control (`SRR7877884 `_); this original dataset is ~4 GB. For testing purposes and for the following examples, we used a 10% sub-sampling of the above dataset: (`SRR7877884-int-0.1.fastq.gz `_). This dataset is already interleaved. @@ -72,42 +74,68 @@ Input A JSON file containing the following information: -1. the path to the input FASTQ file (Illumina paired-end interleaved FASTQ) (recommended the output of the Reads QC workflow.) -2. the contig prefix for the FASTA header -3. the output path -4. input_interleaved (boolean) -5. forwards reads fastq file (required value when input_interleaved is false, otherwise use [] ) -6. reverse reads fastq file (required value when input_interleaved is false, otherwise use [] ) -7. memory (optional) ex: “jgi_metaASM.memory”: “105G” -8. threads (optional) ex: “jgi_metaASM.threads”: “16” +1. the path to the input FASTQ file (Illumina paired-end interleaved FASTQ or PacBio paired-end intereleaved FASTQ) (recommended the output of the Reads QC workflow.) +2. project name: nmdc:XXXXXX +3. memory (optional) ex: "jgi_metaAssembly.memory": "105G" +4. threads (optional) ex: "jgi_metaAssembly.threads": "16" +5. whether the input is short reads (boolean) + +An example input JSON file is shown below for short reads:: + + { + "jgi_metaAssembly.input_files":["https://portal.nersc.gov/project/m3408//test_data/smalltest.int.fastq.gz"], + "jgi_metaAssembly.proj":"nmdc:XXXXXX", + "jgi_metaAssembly.memory": "105G", + "jgi_metaAssembly.threads": "16", + "jgi_metaAssembly.shortRead": true + } + -An example input JSON file is shown below:: +An example input JSON file is shown below for long reads:: { - "jgi_metaASM.input_file":["/path/to/SRR7877884-int-0.1.fastq.gz "], - "jgi_metaASM.rename_contig_prefix":"projectID", - "jgi_metaASM.outdir":"/path/to/ SRR7877884-int-0.1_assembly", - "jgi_metaASM.input_interleaved":true, - "jgi_metaASM.input_fq1":[], - "jgi_metaASM.input_fq2":[], - "jgi_metaASM.memory": "105G", - "jgi_metaASM.threads": "16" + "jgi_metaAssembly.input_files":["/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz"], + "jgi_metaAssembly.proj":"nmdc:XXXXXX", + "jgi_metaAssembly.memory": "105G", + "jgi_metaAssembly.threads": "16", + "jgi_metaAssembly.shortRead": false } Output ------ -The output directory will contain following files:: +The output directory will contain following files for short reads:: output/ - ├── assembly.agp - ├── assembly_contigs.fna - ├── assembly_scaffolds.fna - ├── covstats.txt - ├── pairedMapped.sam.gz - ├── pairedMapped_sorted.bam - └── stats.json + ├── nmdc_XXXXXX_metaAsm.info + ├── nmdc_XXXXXX_covstats.txt + ├── nmdc_XXXXXX_bbcms.fastq.gz + ├── nmdc_XXXXXX_scaffolds.fna + ├── nmdc_XXXXXX_assembly.agp + ├── stats.json + ├── nmdc_XXXXXX_pairedMapped.sam.gz + └── nmdc_XXXXXX_pairedMapped_sorted.bam + + +The output directory will contain following files for long reads:: + + + output/ + ├── nmdc_XXXXXX_assembly.legend + ├── nmdc_XXXXXX_contigs.fna + ├── nmdc_XXXXXX_pairedMapped_sorted.bam + ├── nmdc_XXXXXX_read_count_report.txt + ├── nmdc_XXXXXX_metaAsm.info + ├── nmdc_XXXXXX__summary.stats + ├── nmdc_XXXXXX_scaffolds.fna + ├── nmdc_XXXXXX_pairedMapped.sam.gz + ├── stats.json + ├── nmdc_XXXXXX_contigs.sam.stats + ├── nmdc_XXXXXX_contigs.sorted.bam.pileup.basecov + ├── nmdc_XXXXXX_assembly.agp + └── nmdc_XXXXXX_contigs.sorted.bam.pileup.out + Part of an example output stats JSON file is shown below: @@ -137,8 +165,7 @@ Part of an example output stats JSON file is shown below: "scaf_l_gt50K": 0, "scaf_pct_gt50K": 0.0, "gc_avg": 0.39129, - "gc_std": 0.03033, - "filename": "/global/cfs/cdirs/m3408/aim2/metagenome/assembly/cromwell-executions/jgi_metaASM/3342a6e8-7f78-40e6-a831-364dd2a47baa/call-create_agp/execution/assembly_scaffolds.fna" + "gc_std": 0.03033 } ``` @@ -224,7 +251,7 @@ mapping/ stdout.background Version History --------------- -- 1.0.2 (release date **03/12/2021**; previous versions: 1.0.1) +- 1.0.6 (release date **11/12/24**; previous versions: 1.0.5) Point of contact ---------------- From df5b86ad7512bbd31154f75184f4551c64d0080e Mon Sep 17 00:00:00 2001 From: Valerie Li Date: Tue, 12 Nov 2024 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From f47f9655d6e14af98fd185463880bde573677711 Mon Sep 17 00:00:00 2001 From: Valerie Li <147084799+vlilanl@users.noreply.github.com> Date: Tue, 12 Nov 2024 09:05:20 -0700 Subject: [PATCH 09/13] Update version.txt --- version.txt | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/version.txt b/version.txt index 7969702..2148fba 100644 --- a/version.txt +++ b/version.txt @@ -1,3 +1,3 @@ -v1.0.6 +v1.0.7 From 86de9345e1b8671d115acf51de5f1625ef875c48 Mon Sep 17 00:00:00 2001 From: Valerie Li <147084799+vlilanl@users.noreply.github.com> Date: Tue, 12 Nov 2024 09:05:40 -0700 Subject: [PATCH 10/13] Update index.rst --- docs/index.rst | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index ba0becf..fdcb751 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -1,4 +1,4 @@ -Metagenome Assembly Workflow (v1.0.6) +Metagenome Assembly Workflow (v1.0.7) ======================================== .. image:: workflow_assembly.png @@ -251,7 +251,7 @@ mapping/ stdout.background Version History --------------- -- 1.0.6 (release date **11/12/24**; previous versions: 1.0.5) +- 1.0.7 (release date **11/12/24**; previous versions: 1.0.6) Point of contact ---------------- From 0f4754946732464211b86967325d2f832e72e4cf Mon Sep 17 00:00:00 2001 From: Valerie Li <147084799+vlilanl@users.noreply.github.com> Date: Tue, 12 Nov 2024 09:23:53 -0700 Subject: [PATCH 11/13] Update index.rst --- docs/index.rst | 140 ++++++++++++++++++++++++------------------------- 1 file changed, 68 insertions(+), 72 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index fdcb751..2a73807 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -1,26 +1,29 @@ Metagenome Assembly Workflow (v1.0.7) -======================================== +===================================== .. image:: workflow_assembly.png :scale: 60% :alt: Metagenome assembly workflow dependencies - + Workflow Overview ----------------- -This workflow takes in paired-end Illumina short reads or paired-end PacBio long reads in interleaved format and performs error correction, then reformats the interleaved file into two FASTQ files for downstream tasks using bbcms (BBTools). The corrected reads are assembled using metaSPAdes. After assembly, the reads are mapped back to contigs by bbmap (BBTools) for coverage information. The .wdl (Workflow Description Language) file includes five tasks, *bbcms*, *assy*, *create_agp*, *read_mapping_pairs*, and *make_output*. +This workflow takes in paired-end Illumina short reads or paired-end PacBio long reads in interleaved format and performs error correction, then reformats the interleaved file into two FASTQ files for downstream tasks using bbcms (BBTools). The corrected reads are assembled using metaSPAdes. After assembly, the reads are mapped back to contigs by bbmap (BBTools) for coverage information. The `.wdl` (Workflow Description Language) file includes five tasks: *bbcms*, *assy*, *create_agp*, *read_mapping_pairs*, and *make_output*. -1. The *bbcms* task takes in interleaved FASTQ inputs and performs error correction and reformats the interleaved fastq into two output FASTQ files for paired-end reads for the next tasks. -2. The *assy* task performs metaSPAdes assembly -3. Contigs and Scaffolds (output of metaSPAdes) are consumed by the *create_agp* task to rename the FASTA header and generate an `AGP format `_ which describes the assembly +1. The *bbcms* task takes in interleaved FASTQ inputs, performs error correction, and reformats the interleaved FASTQ into two output FASTQ files for paired-end reads for the next tasks. +2. The *assy* task performs metaSPAdes assembly. +3. Contigs and Scaffolds (output of metaSPAdes) are processed by the *create_agp* task to rename the FASTA header and generate an `AGP format `_ which describes the assembly. 4. The *read_mapping_pairs* task maps reads back to the final assembly to generate coverage information. -5. The final *make_output* task adds all output files into the specified directory. +5. The final *make_output* task collects all output files into the specified directory. Workflow Availability --------------------- -The workflow from GitHub uses all the listed docker images to run all third-party tools. -The workflow is available in GitHub: https://github.com/microbiomedata/metaAssembly; the corresponding Docker images are available in DockerHub: https://hub.docker.com/r/microbiomedata/spades and https://hub.docker.com/r/microbiomedata/bbtools +The workflow from GitHub uses all the listed Docker images to run all third-party tools. +The workflow is available on GitHub: `https://github.com/microbiomedata/metaAssembly` +The corresponding Docker images are available on DockerHub: +- `https://hub.docker.com/r/microbiomedata/spades` +- `https://hub.docker.com/r/microbiomedata/bbtools` Requirements for Execution -------------------------- @@ -40,62 +43,61 @@ The memory requirement depends on the input complexity. Here is a simple estimat predicted_mem = (kmers * 2.962e-08 + 1.630e+01) * 1.1 (in GB) .. note:: - - The kmers variable for the equation above can be obtained using the kmercountmulti.sh script from BBTools. - kmercountmulti.sh -k=31 in=your.read.fq.gz + The kmers variable for the equation above can be obtained using the `kmercountmulti.sh` script from BBTools. + + Example command: + + :: + kmercountmulti.sh -k=31 in=your.read.fq.gz Workflow Dependencies --------------------- -Third party software: (This is included in the Docker image.) +Third-party software: (This is included in the Docker image.) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -- `metaSPades v3.15.0 `_ (License: `GPLv2 `_) -- `BBTools:38.94 `_ (License: `BSD-3-Clause-LBNL `_) +- `metaSPAdes v3.15.0 `_ (License: `GPLv2 `_) +- `BBTools v38.94 `_ (License: `BSD-3-Clause-LBNL `_) Sample dataset(s) ----------------- -- small dataset: `Ecoli 10x (287M) `_ . You can find input/output in the downloaded tar gz file. - -- large dataset: `Zymobiomics mock-community DNA control (22G) `_ . You can find input/output in the downloaded tar gz file. - -- long reads dataset: `/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz` - -Zymobiomics mock-community DNA control (`SRR7877884 `_); this original dataset is ~4 GB. +- Small dataset: `Ecoli 10x (287M) `_ (Input/output included in tar.gz file) +- Large dataset: `Zymobiomics mock-community DNA control (22G) `_ (Input/output included in tar.gz file) +- Long reads dataset: `PacBio `_ -For testing purposes and for the following examples, we used a 10% sub-sampling of the above dataset: (`SRR7877884-int-0.1.fastq.gz `_). This dataset is already interleaved. +Zymobiomics mock-community DNA control (`SRR7877884 `_). The original dataset is ~4 GB. +For testing, a 10% subsample of the dataset is used: (`SRR7877884-int-0.1.fastq.gz `_). This dataset is already interleaved. Input ----- A JSON file containing the following information: -1. the path to the input FASTQ file (Illumina paired-end interleaved FASTQ or PacBio paired-end intereleaved FASTQ) (recommended the output of the Reads QC workflow.) -2. project name: nmdc:XXXXXX -3. memory (optional) ex: "jgi_metaAssembly.memory": "105G" -4. threads (optional) ex: "jgi_metaAssembly.threads": "16" -5. whether the input is short reads (boolean) +1. The path to the input FASTQ file (Illumina paired-end interleaved FASTQ or PacBio paired-end interleaved FASTQ) (recommended: output of the Reads QC workflow). +2. Project name: nmdc:XXXXXX +3. Memory (optional) e.g., `"jgi_metaAssembly.memory": "105G"` +4. Threads (optional) e.g., `"jgi_metaAssembly.threads": "16"` +5. Whether the input is short reads (boolean) -An example input JSON file is shown below for short reads:: +Example input JSON for short reads:: { - "jgi_metaAssembly.input_files":["https://portal.nersc.gov/project/m3408//test_data/smalltest.int.fastq.gz"], - "jgi_metaAssembly.proj":"nmdc:XXXXXX", + "jgi_metaAssembly.input_files": ["https://portal.nersc.gov/project/m3408/test_data/smalltest.int.fastq.gz"], + "jgi_metaAssembly.proj": "nmdc:XXXXXX", "jgi_metaAssembly.memory": "105G", "jgi_metaAssembly.threads": "16", "jgi_metaAssembly.shortRead": true } - -An example input JSON file is shown below for long reads:: +Example input JSON for long reads:: { - "jgi_metaAssembly.input_files":["/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz"], - "jgi_metaAssembly.proj":"nmdc:XXXXXX", + "jgi_metaAssembly.input_files": ["/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz"], + "jgi_metaAssembly.proj": "nmdc:XXXXXX", "jgi_metaAssembly.memory": "105G", "jgi_metaAssembly.threads": "16", "jgi_metaAssembly.shortRead": false @@ -104,8 +106,7 @@ An example input JSON file is shown below for long reads:: Output ------ -The output directory will contain following files for short reads:: - +The output directory will contain the following files for short reads:: output/ ├── nmdc_XXXXXX_metaAsm.info @@ -117,9 +118,7 @@ The output directory will contain following files for short reads:: ├── nmdc_XXXXXX_pairedMapped.sam.gz └── nmdc_XXXXXX_pairedMapped_sorted.bam - -The output directory will contain following files for long reads:: - +The output directory will contain the following files for long reads:: output/ ├── nmdc_XXXXXX_assembly.legend @@ -127,7 +126,7 @@ The output directory will contain following files for long reads:: ├── nmdc_XXXXXX_pairedMapped_sorted.bam ├── nmdc_XXXXXX_read_count_report.txt ├── nmdc_XXXXXX_metaAsm.info - ├── nmdc_XXXXXX__summary.stats + ├── nmdc_XXXXXX_summary.stats ├── nmdc_XXXXXX_scaffolds.fna ├── nmdc_XXXXXX_pairedMapped.sam.gz ├── stats.json @@ -136,38 +135,35 @@ The output directory will contain following files for long reads:: ├── nmdc_XXXXXX_assembly.agp └── nmdc_XXXXXX_contigs.sorted.bam.pileup.out +Example output stats JSON file:: -Part of an example output stats JSON file is shown below: - -``` -{ - "scaffolds": 58, - "contigs": 58, - "scaf_bp": 28406, - "contig_bp": 28406, - "gap_pct": 0.00000, - "scaf_N50": 21, - "scaf_L50": 536, - "ctg_N50": 21, - "ctg_L50": 536, - "scaf_N90": 49, - "scaf_L90": 317, - "ctg_N90": 49, - "ctg_L90": 317, - "scaf_logsum": 22.158, - "scaf_powsum": 2.245, - "ctg_logsum": 22.158, - "ctg_powsum": 2.245, - "asm_score": 0.000, - "scaf_max": 1117, - "ctg_max": 1117, - "scaf_n_gt50K": 0, - "scaf_l_gt50K": 0, - "scaf_pct_gt50K": 0.0, - "gc_avg": 0.39129, - "gc_std": 0.03033 -} -``` + { + "scaffolds": 58, + "contigs": 58, + "scaf_bp": 28406, + "contig_bp": 28406, + "gap_pct": 0.00000, + "scaf_N50": 21, + "scaf_L50": 536, + "ctg_N50": 21, + "ctg_L50": 536, + "scaf_N90": 49, + "scaf_L90": 317, + "ctg_N90": 49, + "ctg_L90": 317, + "scaf_logsum": 22.158, + "scaf_powsum": 2.245, + "ctg_logsum": 22.158, + "ctg_powsum": 2.245, + "asm_score": 0.000, + "scaf_max": 1117, + "ctg_max": 1117, + "scaf_n_gt50K": 0, + "scaf_l_gt50K": 0, + "scaf_pct_gt50K": 0.0, + "gc_avg": 0.39129, + "gc_std": 0.03033 + } The table provides all of the output directories, files, and their descriptions. From 4676bf3dfbc68753dc5b6d5b01a397ea4d16c0bb Mon Sep 17 00:00:00 2001 From: Valerie Li <147084799+vlilanl@users.noreply.github.com> Date: Tue, 12 Nov 2024 09:35:12 -0700 Subject: [PATCH 12/13] Update index.rst --- docs/index.rst | 10 +++++++--- 1 file changed, 7 insertions(+), 3 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 2a73807..4bb6a47 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -20,8 +20,11 @@ Workflow Availability --------------------- The workflow from GitHub uses all the listed Docker images to run all third-party tools. + The workflow is available on GitHub: `https://github.com/microbiomedata/metaAssembly` + The corresponding Docker images are available on DockerHub: + - `https://hub.docker.com/r/microbiomedata/spades` - `https://hub.docker.com/r/microbiomedata/bbtools` @@ -36,7 +39,7 @@ Requirements for Execution Hardware Requirements --------------------- -- Memory: >40 GB RAM +**Memory: >40 GB RAM** The memory requirement depends on the input complexity. Here is a simple estimation equation for the memory required based on kmers in the input file:: @@ -66,7 +69,8 @@ Sample dataset(s) - Small dataset: `Ecoli 10x (287M) `_ (Input/output included in tar.gz file) - Large dataset: `Zymobiomics mock-community DNA control (22G) `_ (Input/output included in tar.gz file) -- Long reads dataset: `PacBio `_ + +Long reads dataset: `PacBio `_ Zymobiomics mock-community DNA control (`SRR7877884 `_). The original dataset is ~4 GB. @@ -75,7 +79,7 @@ For testing, a 10% subsample of the dataset is used: (`SRR7877884-int-0.1.fastq. Input ----- -A JSON file containing the following information: +A `JSON file `_ containing the following information: 1. The path to the input FASTQ file (Illumina paired-end interleaved FASTQ or PacBio paired-end interleaved FASTQ) (recommended: output of the Reads QC workflow). 2. Project name: nmdc:XXXXXX From 34e83d1bfff89e142ae7cb45dc31ada411e14e27 Mon Sep 17 00:00:00 2001 From: Valerie Li <147084799+vlilanl@users.noreply.github.com> Date: Thu, 14 Nov 2024 15:23:02 -0700 Subject: [PATCH 13/13] Revert index.rst --- docs/index.rst | 189 +++++++++++++++++++++---------------------------- 1 file changed, 81 insertions(+), 108 deletions(-) diff --git a/docs/index.rst b/docs/index.rst index 4bb6a47..589b96f 100644 --- a/docs/index.rst +++ b/docs/index.rst @@ -1,32 +1,26 @@ -Metagenome Assembly Workflow (v1.0.7) +Metagenome Assembly Workflow (v1.0.2) ===================================== .. image:: workflow_assembly.png :scale: 60% :alt: Metagenome assembly workflow dependencies - + Workflow Overview ----------------- -This workflow takes in paired-end Illumina short reads or paired-end PacBio long reads in interleaved format and performs error correction, then reformats the interleaved file into two FASTQ files for downstream tasks using bbcms (BBTools). The corrected reads are assembled using metaSPAdes. After assembly, the reads are mapped back to contigs by bbmap (BBTools) for coverage information. The `.wdl` (Workflow Description Language) file includes five tasks: *bbcms*, *assy*, *create_agp*, *read_mapping_pairs*, and *make_output*. +This workflow takes in paired-end Illumina reads in interleaved format and performs error correction, then reformats the interleaved file into two FASTQ files for downstream tasks using bbcms (BBTools). The corrected reads are assembled using metaSPAdes. After assembly, the reads are mapped back to contigs by bbmap (BBTools) for coverage information. The .wdl (Workflow Description Language) file includes five tasks, *bbcms*, *assy*, *create_agp*, *read_mapping_pairs*, and *make_output*. -1. The *bbcms* task takes in interleaved FASTQ inputs, performs error correction, and reformats the interleaved FASTQ into two output FASTQ files for paired-end reads for the next tasks. -2. The *assy* task performs metaSPAdes assembly. -3. Contigs and Scaffolds (output of metaSPAdes) are processed by the *create_agp* task to rename the FASTA header and generate an `AGP format `_ which describes the assembly. +1. The *bbcms* task takes in interleaved FASTQ inputs and performs error correction and reformats the interleaved fastq into two output FASTQ files for paired-end reads for the next tasks. +2. The *assy* task performs metaSPAdes assembly +3. Contigs and Scaffolds (output of metaSPAdes) are consumed by the *create_agp* task to rename the FASTA header and generate an `AGP format `_ which describes the assembly 4. The *read_mapping_pairs* task maps reads back to the final assembly to generate coverage information. -5. The final *make_output* task collects all output files into the specified directory. +5. The final *make_output* task adds all output files into the specified directory. Workflow Availability --------------------- -The workflow from GitHub uses all the listed Docker images to run all third-party tools. - -The workflow is available on GitHub: `https://github.com/microbiomedata/metaAssembly` - -The corresponding Docker images are available on DockerHub: - -- `https://hub.docker.com/r/microbiomedata/spades` -- `https://hub.docker.com/r/microbiomedata/bbtools` +The workflow from GitHub uses all the listed docker images to run all third-party tools. +The workflow is available in GitHub: https://github.com/microbiomedata/metaAssembly; the corresponding Docker images are available in DockerHub: https://hub.docker.com/r/microbiomedata/spades and https://hub.docker.com/r/microbiomedata/bbtools Requirements for Execution -------------------------- @@ -39,135 +33,114 @@ Requirements for Execution Hardware Requirements --------------------- -**Memory: >40 GB RAM** +- Memory: >40 GB RAM The memory requirement depends on the input complexity. Here is a simple estimation equation for the memory required based on kmers in the input file:: predicted_mem = (kmers * 2.962e-08 + 1.630e+01) * 1.1 (in GB) .. note:: + + The kmers variable for the equation above can be obtained using the kmercountmulti.sh script from BBTools. - The kmers variable for the equation above can be obtained using the `kmercountmulti.sh` script from BBTools. + kmercountmulti.sh -k=31 in=your.read.fq.gz - Example command: - - :: - - kmercountmulti.sh -k=31 in=your.read.fq.gz Workflow Dependencies --------------------- -Third-party software: (This is included in the Docker image.) +Third party software: (This is included in the Docker image.) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -- `metaSPAdes v3.15.0 `_ (License: `GPLv2 `_) -- `BBTools v38.94 `_ (License: `BSD-3-Clause-LBNL `_) +- `metaSPades v3.15.0 `_ (License: `GPLv2 `_) +- `BBTools:38.94 `_ (License: `BSD-3-Clause-LBNL `_) Sample dataset(s) ----------------- -- Small dataset: `Ecoli 10x (287M) `_ (Input/output included in tar.gz file) -- Large dataset: `Zymobiomics mock-community DNA control (22G) `_ (Input/output included in tar.gz file) +- small dataset: `Ecoli 10x (287M) `_ . You can find input/output in the downloaded tar gz file. -Long reads dataset: `PacBio `_ +- large dataset: `Zymobiomics mock-community DNA control (22G) `_ . You can find input/output in the downloaded tar gz file. -Zymobiomics mock-community DNA control (`SRR7877884 `_). The original dataset is ~4 GB. +Zymobiomics mock-community DNA control (`SRR7877884 `_); this original dataset is ~4 GB. + +For testing purposes and for the following examples, we used a 10% sub-sampling of the above dataset: (`SRR7877884-int-0.1.fastq.gz `_). This dataset is already interleaved. -For testing, a 10% subsample of the dataset is used: (`SRR7877884-int-0.1.fastq.gz `_). This dataset is already interleaved. Input ----- -A `JSON file `_ containing the following information: - -1. The path to the input FASTQ file (Illumina paired-end interleaved FASTQ or PacBio paired-end interleaved FASTQ) (recommended: output of the Reads QC workflow). -2. Project name: nmdc:XXXXXX -3. Memory (optional) e.g., `"jgi_metaAssembly.memory": "105G"` -4. Threads (optional) e.g., `"jgi_metaAssembly.threads": "16"` -5. Whether the input is short reads (boolean) +A JSON file containing the following information: -Example input JSON for short reads:: +1. the path to the input FASTQ file (Illumina paired-end interleaved FASTQ) (recommended the output of the Reads QC workflow.) +2. the contig prefix for the FASTA header +3. the output path +4. input_interleaved (boolean) +5. forwards reads fastq file (required value when input_interleaved is false, otherwise use [] ) +6. reverse reads fastq file (required value when input_interleaved is false, otherwise use [] ) +7. memory (optional) ex: “jgi_metaASM.memory”: “105G” +8. threads (optional) ex: “jgi_metaASM.threads”: “16” - { - "jgi_metaAssembly.input_files": ["https://portal.nersc.gov/project/m3408/test_data/smalltest.int.fastq.gz"], - "jgi_metaAssembly.proj": "nmdc:XXXXXX", - "jgi_metaAssembly.memory": "105G", - "jgi_metaAssembly.threads": "16", - "jgi_metaAssembly.shortRead": true - } - -Example input JSON for long reads:: +An example input JSON file is shown below:: { - "jgi_metaAssembly.input_files": ["/global/cfs/cdirs/m3408/www/test_data/SRR13128014.pacbio.subsample.ccs.fastq.gz"], - "jgi_metaAssembly.proj": "nmdc:XXXXXX", - "jgi_metaAssembly.memory": "105G", - "jgi_metaAssembly.threads": "16", - "jgi_metaAssembly.shortRead": false + "jgi_metaASM.input_file":["/path/to/SRR7877884-int-0.1.fastq.gz "], + "jgi_metaASM.rename_contig_prefix":"projectID", + "jgi_metaASM.outdir":"/path/to/ SRR7877884-int-0.1_assembly", + "jgi_metaASM.input_interleaved":true, + "jgi_metaASM.input_fq1":[], + "jgi_metaASM.input_fq2":[], + "jgi_metaASM.memory": "105G", + "jgi_metaASM.threads": "16" } Output ------ -The output directory will contain the following files for short reads:: - - output/ - ├── nmdc_XXXXXX_metaAsm.info - ├── nmdc_XXXXXX_covstats.txt - ├── nmdc_XXXXXX_bbcms.fastq.gz - ├── nmdc_XXXXXX_scaffolds.fna - ├── nmdc_XXXXXX_assembly.agp - ├── stats.json - ├── nmdc_XXXXXX_pairedMapped.sam.gz - └── nmdc_XXXXXX_pairedMapped_sorted.bam +The output directory will contain following files:: -The output directory will contain the following files for long reads:: output/ - ├── nmdc_XXXXXX_assembly.legend - ├── nmdc_XXXXXX_contigs.fna - ├── nmdc_XXXXXX_pairedMapped_sorted.bam - ├── nmdc_XXXXXX_read_count_report.txt - ├── nmdc_XXXXXX_metaAsm.info - ├── nmdc_XXXXXX_summary.stats - ├── nmdc_XXXXXX_scaffolds.fna - ├── nmdc_XXXXXX_pairedMapped.sam.gz - ├── stats.json - ├── nmdc_XXXXXX_contigs.sam.stats - ├── nmdc_XXXXXX_contigs.sorted.bam.pileup.basecov - ├── nmdc_XXXXXX_assembly.agp - └── nmdc_XXXXXX_contigs.sorted.bam.pileup.out - -Example output stats JSON file:: - - { - "scaffolds": 58, - "contigs": 58, - "scaf_bp": 28406, - "contig_bp": 28406, - "gap_pct": 0.00000, - "scaf_N50": 21, - "scaf_L50": 536, - "ctg_N50": 21, - "ctg_L50": 536, - "scaf_N90": 49, - "scaf_L90": 317, - "ctg_N90": 49, - "ctg_L90": 317, - "scaf_logsum": 22.158, - "scaf_powsum": 2.245, - "ctg_logsum": 22.158, - "ctg_powsum": 2.245, - "asm_score": 0.000, - "scaf_max": 1117, - "ctg_max": 1117, - "scaf_n_gt50K": 0, - "scaf_l_gt50K": 0, - "scaf_pct_gt50K": 0.0, - "gc_avg": 0.39129, - "gc_std": 0.03033 - } + ├── assembly.agp + ├── assembly_contigs.fna + ├── assembly_scaffolds.fna + ├── covstats.txt + ├── pairedMapped.sam.gz + ├── pairedMapped_sorted.bam + └── stats.json + +Part of an example output stats JSON file is shown below: + +``` +{ + "scaffolds": 58, + "contigs": 58, + "scaf_bp": 28406, + "contig_bp": 28406, + "gap_pct": 0.00000, + "scaf_N50": 21, + "scaf_L50": 536, + "ctg_N50": 21, + "ctg_L50": 536, + "scaf_N90": 49, + "scaf_L90": 317, + "ctg_N90": 49, + "ctg_L90": 317, + "scaf_logsum": 22.158, + "scaf_powsum": 2.245, + "ctg_logsum": 22.158, + "ctg_powsum": 2.245, + "asm_score": 0.000, + "scaf_max": 1117, + "ctg_max": 1117, + "scaf_n_gt50K": 0, + "scaf_l_gt50K": 0, + "scaf_pct_gt50K": 0.0, + "gc_avg": 0.39129, + "gc_std": 0.03033, + "filename": "/global/cfs/cdirs/m3408/aim2/metagenome/assembly/cromwell-executions/jgi_metaASM/3342a6e8-7f78-40e6-a831-364dd2a47baa/call-create_agp/execution/assembly_scaffolds.fna" +} +``` The table provides all of the output directories, files, and their descriptions. @@ -251,7 +224,7 @@ mapping/ stdout.background Version History --------------- -- 1.0.7 (release date **11/12/24**; previous versions: 1.0.6) +- 1.0.2 (release date **03/12/2021**; previous versions: 1.0.1) Point of contact ----------------

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