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
