Help me! / Yeast transformation in Two-Hybrid screening

Takashi Nagano naganot at bri.niigata-u.ac.jp
Mon Aug 19 04:22:53 EST 1996


I am now doing Two-Hybrid screening with CLONTECH's MATCHMAKER 2 system
(yeast: CG1945, bait plasmid: pAS2-1) and pGAD10-based library. My
problem is too low transformation efficiency in sequential
transformation. I have tried on several protocols including one in
CLONTECH's manual, but the efficiency is no more than 4000cfu/microgram
plasmid so far. I hear that 200 thousand cfu/microgram plasmid can be
achieved by some researchers. Someone who knows an optimal or better
protocol and/or SD medium recipe for transforming CG1945 yeast, please
teach me how they are!
Thank you very much for your help. 

My e-mail address is : naganot at bri.niigata-u.ac.jp

Takashi Nagano



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