2-hybrid sys

Kathy Shire shirek at FHS.CSU.MCMASTER.CA
Tue Sep 13 14:59:12 EST 1994


Hi guys!

There was a problem (a while ago) about screening a library and getting a
redundant reduntant clone (was it gal4?).

The question was "is there a way to pre-screen clones?" 

Recently, I have experienced the same problem with a different, out of
frame, protein.  There is a way to screen your clones before doing large
scale preps and sequencing.  Do a restriction analysis of the evil piece of
DNA and digest small scale DH5a (shuttled from yeast).  I have managed to
get 4 clones (thank God!) out of 150 that don't resemble the out-of-frame
protein without a lot of extra work.

I hope this will help in some way.

You're not alone...

Kath








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