BclI

Darren Natale camdna at ubvmsb.cc.buffalo.edu
Wed Sep 16 16:52:00 EST 1992


In article <1992Sep16.163757.22921 at murdoch.acc.Virginia.EDU>, mdv6n at dayhoff.med.Virginia.EDU (Mark D. Van Cleve) writes...
> 
> 
>Anyone else have trouble with this enzyme?  Either it goes bad very quickly
>in our freezer, which would be odd for a heat stable enzyme, or we're not
>using it correctly, or something, because BclI from several different 
>sources has very little interest in cutting plasmid for us, even when it
>will consent to cut lambda, which is not always.  Any clues out there?>
> 

Bcl I is sensitive to dam methylation (methylation of A bases in the sequence
GATC). Since this enzyme's recognition site contains a dam site as well, any 
plasmid, etc. isolated from dam+ bacteria should be (in theory) entirely
resistant to Bcl I digestion. Try isolating your DNA from a dam- strain.

DN



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