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2-hybrid no blue his+ clones

ehrhardt at mpimg-berlin-dahlem.mpg.de ehrhardt at mpimg-berlin-dahlem.mpg.de
Wed Nov 15 05:19:02 EST 1995


Dear netters...

I am glad to hear some of you doing fine with lacZ assay in 2-Hybrid
system. My problem is just the opposite. I did a 2 Hy. screen of 500.000
clones using Gal4 system and found about 80 positive (i.e. His+) clones.
checking them in lacZ assay I failed to detect blue clones at all. My controls
(p53, pSV40 interacting controls from Stratagene) do work fine. In another
screen (different bait) I found only one of 12 His+ clones turning blue in the
assay. 
My question: Did someone have similar problems? Any idea why this may happen?

Thanks,
Tom



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