Genomic library in pZErO

David L. Haviland, Ph.D. dhavilan at IMM2.IMM.UTH.TMC.EDU
Tue Dec 8 12:58:57 EST 1998


At 09:15 12/8/98 -0800, bbeitzel at biomail.ucsd.edu wrote:

Brett:

>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.

Thanks this is good to know!

>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.

They do sell a pZERO kan or at least they did.  I bought it well over a
year ago and frankly had very little luck with it.  Granted, I only tried
to use it twice and all I managed to get back was just vector.  The inserts
I tried ranged from 1.2 kb all the way up to 16 kb.

I too wanted the kan rather than zeocin.

David



More information about the Methods mailing list