Genomic library in pZErO

bbeitzel at biomail.ucsd.edu bbeitzel at biomail.ucsd.edu
Tue Dec 8 12:15:28 EST 1998


I have found that I usually have to CIP the pZErO vectors when working
with large inserts.  They do have reduced background, but it is possible
to generate vector multimers that do not express ccdB.  This becomes more
apparent when you are using very large inserts, in which case the vector
multimers are probably the favored ligation product.
I have also found the Zeocin is a pain to work with.  You usually need to
make fresh plates 1-2 days before you need them, because even after only
one week in the cold room the zeocin starts to go off.  I have made an amp
resistant derivative of pZErO to avoid using zeocin, and I think
Invitrogen now sells a kan version.

Hope some of this helps.

Cheers,
Brett Beitzel
bbeitzel at biomail.ucsd.edu

Infectious Disease Lab
The Salk Institute
La Jolla, CA 


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