[Plant-biology] pCambia1201 question

James J. Campanella via plantbio%40net.bio.net (by campanellj from mail.montclair.edu)
Tue Jul 1 18:34:50 EST 2008


Dear fellow workers in the mines of science,

I have been tryingto work with the pCambia1201 plasmid for the lastseveral weeks, but ithas been driving my lab a bit crazy. My problemis that it does not wantto digest with most endonucleases we've tried.I simply can not use theplasmid if I can't cut it, so I have aproblem. It was suggested to meto try transferring the plasmid to DH5cells, which I did to no avail--the resultant plasmid still does notcut with almost every enzyme in mylibrary. I was finally able to getit to cut with XhoI and PvuII, buteven those were only partialdigests.

Does anybody have anyadvice on working with this thing? The literaturedoes not suggest thatthere is even a problem, and the group thatengineered it could onlysuggest "trying all your enzymes until youfind one that cuts", which isa bit impractical.

Thanks for any help,

Jim Campanella

-------------------

James J. Campanella,
Associate Professor,
Department of Biology and Molecular Biology
Montclair State University
1 Normal Avenue
Montclair, NJ 07043

Alternate email address: jcamp from alumni.uchicago.edu

Ph: 973-655-4097
Fax: 973-655-7047



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