Heyer Lab

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Protocols

– Lab jobs

– Yeast protocols

  • Basics on Saccharomyces cerevisiae
  • General tips
  • DNA extraction using CTAB
  • DNA extraction using PCIA
  • DNA extraction in agarose plugs (for Pulse-Field gel)
  • DNA extraction using KAc and Isopropanol
  • Gene expression induction with Galactose (like the HO endonuclease with the GAL-HO construct)
  • Immunoprecipitation
  • Irradiation (gamma-ray)
  • Media recipes
  • MMS handling
  • PCR on colony (fast transformant screening)
  • Plasmid transfer from E. coli to S. cerevisiae
  • Proteins extraction using TCA
  • Psoralen crosslinking and DNA extraction
  • Psoralen crosslinking (variation very severe crosslinking density)
  • Psoralen de-crosslinking
  • Yeast restric
  • Yeast sporulation
  • Yeast spotting assay
  • Yeast strains archiving
  • Yeast transformation / Fast yeast transformation

– Biochemistry protocols

  • DNA strand exchange reaction (D-loop assay)
  • ATPase assay
  • Nucleoprotein gel assay for Rad51-DNA complexes
  • Fermentor protocol

– Molecular biology various

  • Ligation (sticky ends)
  • Ligation (blunt ends)
  • DNA precipitation with Ethanol and salts
  • Dialysis tube preparation
  • Dephosphorylation of plasmid
  • Denaturing purification of insoluble proteins
  • Lab Solutions

– Gels

  • Antibody production
  • Western blot
  • Southern blot
  • Northern blot
  • 2D-gel
  • Protein polyacrylamide gel
  • DNA polyacrylamide gel
  • Pulse-Field gel
  • PCR
  • Real Time qPCR

– E. coli

  • Ampicillin preparation
  • E. coli plasmid transformation
  • E. coli plasmid extraction
  • E. coli proteins extraction
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