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Herskowitz
Lab Protocol
From : via Caroline Shamu,
Ref : BioTechniques 13:18 (1992)
Sleazy
Plate Transformation of Yeast
- Take 0.5ml
culture, or toothpick full yeast off a plate
- Spin, then
remove sup by inverting and shaking once
- add 10ul carrier
DNA (=100ug),
- add 1-5ug
transforming DNA (more is better)
- vortex
- 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
- vortex
- incubate overnight
on benchtop
- plate directly
by taking bottom layer, or spinning down/resuspending
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