BsaBI digests

Bernard Murray bernard at
Tue Jan 16 12:21:41 EST 1996

In article <JDuPvKAm75+wEw4R at>, duncan at 
>In article <v01530501ad207196f61c@[]>, jlight at RESUNIX.RI.SICK
>KIDS.ON.CA writes
>>Does anyone out there have experience doing BsaBI digests?  It's one of
>>those enzymes with activity that's blocked by overlapping dam methylation.
>>I'm trying to cut a plasmid with a single BsaBI site.  The plasmid was
>>recovered from strain GM2163, which is dam(-) and dcm(-), but the digest
>>doesn't appear to be working any better.  Any advice would be greatly

Just wondering, did you grow the GM2163 in the presence of chloramphenicol
to maintain the methylation negative status?  I have my stocks selected
at 100 mg/l and plasmids with ClaI in a dam context cut to completetion
when this is used as the host (no cut at all with XL-1 or DH5alpha).

>Can you cut the supposed dam- plasmid with Mbo I. If so it is definitely

This would be a good test of plasmid methylation status and your BsaBI.

	Happy cutting,
Bernard Murray, Ph.D.
bernard at  (National Cancer Institute, NIH, Bethesda MD, USA)

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