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Problem with XmnI digestion

SUPER AUSTRALIAN BOTANIST u6069416 at ucsvc.ucs.unimelb.edu.au
Sun Dec 24 10:57:30 EST 1995


Dear netters,

I have great problem using pMal-c2 vector for expression of a plant cDNA.
I tried to digest the pMal vector with XmnI then ligate to the 1.7kb blunt
ended insert, but only got a few transformants which don't contain the insert.
The problem seems to be associated with the XmnI digestion as I also tried to
just re-ligate the linear vector (not dephosphorylated) and again only obtained 
about 10 colonies. The electoporation system I'm using worked very well for
other transformations. So maybe there is something in the XmnI that is
degrading the blunt ends. I'm using XmnI from NEB, has anyone had a similar 
problem? Any suggestions are greatly appreciated!

Du
P.S. please email me at hedu at mbcl.rutgers.edu
 



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