why no insert from blue colonies

Dong Zhang dongzha at yahoo.com
Sun Apr 14 22:53:07 EST 2002

Hi! Everybody:

I need help on yeast two-hybrid library screening.

I got many positive (blue) colonies using clontech
system 3. The colonies look so blue on
-Trp-Leu-His-Ade plates with X-alpha-gal.

I pick up 100 colonies to do colony PCR for amplify
insert on AD-vector.

The problem is that only, only 40% have shown insert.

Why not another 60% percent? Are they false positive?

If there is no AD vector at all, how could these
colonies grow on the selection plate?

Dong Zhang



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