Help Me ! / Yeast transformation for Two-Hybrid screening

hellmuth at embl-heidelberg.de hellmuth at embl-heidelberg.de
Fri Aug 23 14:24:25 EST 1996


In article <naganot-1908962026120001 at molnb03.bri.niigata-u.ac.jp>, naganot at bri.niigata-u.ac.jp (Takashi Nagano) writes:
> 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
Look in Dan Gietz' Yeast Transformation Homepage:
http://www.umanitoba.ca/faculties/medicine/human_genetics/gietz/Trafo.html
Klaus Hellmuth



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