His+ Ade+ Laz - yeast two hybrid colonies

Marco Tsui marcotsui2003 at yahoo.com
Wed Mar 10 09:44:24 EST 2004


Dear all,
 
I am performing a yeast two hybrid screening using the Clonetech Matchmaker 3 system. Three days after library transformation (on -L-T-H plates), hundreds of His+ colonies come up and 20-30 of them are Ade+. But strangely enough, these Ade+ colonies failed to come up with a blue color in the turn bluecolony lift assay using X-gal, although the positive control did turn blue. I heard that ADE2 is a very stringent reporter and I expect those Ade+ would be LacZ+..... So what is the problem? or should I use the MEL1 reporter instead of lacZ? Thank you very much!!
 
Marco Tsui
Baptist Univeristy , H.K.

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