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Two hybrid screens/ Troubleshooting

Michaela Monshausen monshausen at uke.uni-hamburg.de
Fri Oct 29 00:01:34 EST 1999


Hello everybody!
In the moment I have a bit trouble with my work on two hybrid screens. So I
want to know if you can help me with experiences you made in screening
libraries with this technique.
First the transformations of the yeast with the bait vector were without
problems, the rate of transformands was good. Because of that fact, i do not
think that my bait vector is toxic for the yeast. Screens made by my colleague
with another bait but the same solutions made no problems at that time, she
had a good rate of clones on the plates. Screening my bait had a low
efficiency on the control plates and no success on the dropout plates. And
from one day to the other the transformations no longer had success. We work
with the LiAc Method and so changed all the solutions and checked the purity
of the DNA. We also transformed the yeast with other bait vectors, which
before made no trouble, but now do. 
If you have any idea of what happened or if you need further informations just
send me an e-mail.


Thank you for spending time on my problem, 
;-))
AG




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