Protocols library

Wet lab protocols.

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Cloning

• PCR
• Site-directed mutagenesis
• PCR Mutagenesis (QuikChange® II)
• Gibson cloning
• Golden Gate Assembly
• Colony PCR
• Yeast colony PCR
• Competent cells preparation
  • RbCl comp cells preparation
  • Inoue comp cells preparation (tricky, but best comp cells ever)
  • Quick 'n dirty CaCl2 comp cells preparation
  • Less quick 'n less dirty CaCl2 comp cells preparation
  • Electrocompetent cells preparation
• Transformations
  • Chemical transformation
  • Electroporation
  • Yeast chemical transformations
• Yeast plasmid purification
• Zymoprep protocol
• Ethanol precipitation

Protein expression and purification

• Autoinduction protocol
• Common promoter induction levels
• Culturing mammalian cells
• Compiled yeast protein expression methods
• Generic LacI/GalR family protein purification protocol
• Benchtop nickel purification
• Ni regeneration protocol
• TEV protease cleavage protocol
• BirA biotinylation protocol
• Preparing FPLC
• FPLC fraction collector
• Cleaning the Sephacryl Column

Protein characterization

• SDS-PAGE protocol
• Coomassie staining protocol
• Western blotting protocol
• Run 1x Gel: Coomassie staining w/ Western blotting protocol
• Dot blot
• Pulldown protocol
• Circular dichroism
• Biolayer interferometry protocol
• Differential scanning fluorometry protocol
• Bradford assay
• ELISA
• Isothermal titration calorimetry
• Flow cytometry

HDX/MS

• LacI HDX: exchange protocol
• Packing a trap column
• Manual icebox LC/MS protocol for HDX samples
• HDX data analysis workflow
• Analyze intact mass on Compass
• Calibrate MS
• Detailed HDX/MS protocol with the LEAP system
• Quench buffers

Other

• Lyophilizer usage
• Plaque assay
• Autoclave and Glasswash use