Herskowitz Lab Protocol
From : via Caroline Shamu,
Ref : BioTechniques 13:18 (1992)


Sleazy Plate Transformation of Yeast

  1. Take 0.5ml culture, or toothpick full yeast off a plate
  2. Spin, then remove sup by inverting and shaking once
  3. add 10ul carrier DNA (=100ug),
  4. add 1-5ug transforming DNA (more is better)
  5. vortex
  6. Add 0.5ml PEG/LiAC/TE = 40% PEG 4000 ie 90ml sterile 45% PEG
    0.1M LiAc 10ml 1M LiAc
    0.01M Tris 7.5 1ml 1M Tris 7.5
    0.01M EDTA 0.2ml 0.5M EDTA
  7. vortex
  8. incubate overnight on benchtop
  9. plate directly by taking bottom layer, or spinning down/resuspending

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