Genomic DNA from Campylobacter

Jayant S. Aphale japhale at cangene.com
Wed Aug 2 11:44:23 EST 1995


I am having problems digesting chromosomal DNA isolated from three
Campylobacter species.  The genomic DNA was isolated using the Qiagen
genomic DNA kit Cat# 13343 according to the manufacterers instructions.

The O.D. 260:280 ratio is 1.80 plus/minus 0.05 a nd looks intact on an
EtBr agarose gel but will not cut with either BamH1 or EcoRI even after
extended digestion with excess enzyme.

The problem is probably not trace EtOH because I precipitataed the genomic
DNA a second time, spooled, washed 70% EtOH and dried it as long as I
dared.  Still didn't cut.

The problem may be DNA modification or something I am missing.  According
to the NEB catalog neither EcoRI or BamHI are blockered by methylation.  Does 
anyone have experience cutting Campylobacter genomic DNA?

Glenn





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