Freimoser:Protocols:FastYeastTransformation

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== Protocol: Smash & grab yeast DNA miniprep==
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== Protocol: Fast yeast transformation==
#50ul carrier DNA
#50ul carrier DNA

Revision as of 07:11, 17 October 2006


Protocol: Fast yeast transformation

  1. 50ul carrier DNA
  2. scrap cells from plate
  3. 1-2ul DNA-plasmid (up to 30-50 ul)
  4. 240 ul PEG (50%)
  5. 36 ul Li-Ac (1M)
  6. resuspend with blue tip
  7. incubate at 30°C or RT at least 30 min
  8. heat shock: 45°C, 15 min (or 42°C for 20 min)
  9. spin down, resuspend in 100ul H2O and plate everything on corresponding medium

Material:

  • Plasmid: 100 ng-1ug
  • carrier DNA: salmon/herring sperm DNA in H2O, 2mg/ml stock, 500 ul aliquots, heat-inactivated at 95°C for 5-6 min and put on ice before use (has to be done once).
  • 50% poly-ethylen-glycol (PEG) (MW 33-50) solution, filter-sterilized
  • 1M Li-Acetate, autoclaved


Reference

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