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High efficiency yeast trafo

Frederik Boernke boernke at ipk-gatersleben.de
Mon Nov 17 11:36:04 EST 1997

Hi all,

I have been persistently trying to transform SC strain Y190 after the
PEG/LiAC/SS-DNA method according to Gietz et al. for a 2-hybrid screen.
But I've never obtained the high efficiencies which should be possible.
Though I followed the protocol exactly, even a 60x up scale only yields
me about 350 000 transformants, which is not enough to cover the library.
Has anybody experience with this method and ever obtained "promised"
efficiencies? Or did anybody reveal some bottlenecks or particular
critical steps, besides those quoted on the Gietz Trafo-Homepage? Or can
anybody recommend a different method which might be more reproduceably?
Any suggestions will be appreciated.




Frederik Boernke
Research Group of  Molecular Plant Physiologie
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
e-mail: boernke at ipk-gatersleben.de

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