yeast one hybrid system

tthamm at gwdg.de tthamm at gwdg.de
Tue Jun 3 07:02:42 EST 1997


Hi netters!

I`m a german graduate student and currently working on my diploma thesis.
Has anyone out there experience with the matchmaker yeast one hybrid
system?
I have severe problems to clone my oligos (51 bp with Eco RI and Xba I /
(SalI in the case of the vector pLacZi) restriction sites on both ends) in
the provided vectors pHis i, pHis i-1 and pLacZ i.
In addition to the cloning strategy described in the manual I made control
experiments digesting the vectors without ligating the oligos!
Transformation of these controls should reveal no colonies, but not in my
case. Many colonies where visible! This may be due to an uncomplete
digest.
Does anybody know a digestion protocol that works with these vectors?

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