Help Me ! / Yeast transformation for Two-Hybrid screening

S.L. Rennie rennie at bldghsc.lan1.umanitoba.ca
Fri Aug 23 14:54:16 EST 1996


In article <naganot-1908962026120001 at molnb03.bri.niigata-u.ac.jp>,
naganot at bri.niigata-u.ac.jp (Takashi Nagano) wrote:

> 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 is
> achieved by some researchers.
> 
> SOMEONE WHO KNOWS an optimal or better protocol and/or SD medium recipe
> for transforming CG1945 yeast to get higher efficiency, 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,
> Niigata, JAPAN


Hi Takashi
   Every once in a while we re-post Dan's web site, all about transformation
optimization and other yeast things....its a terribly long URL but

http://www.umanitoba.ca/faculties/medicine/human_genetics/gietz/Trafo.html


There is a link there to ask questions as well.

Hope this helps.
Sharon

-- 





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