Genomic DNA preps...

Paul Tadich tadichp at io.org
Sun Aug 11 16:19:14 EST 1996


Hi.

I'm having a bit of trouble making a genomic DNA prep from
Brevibacterium sterolicum. I've tried the boiling method and a
standard phenol extraction, but these two procedures don't seem to be
working very well. My best purification to date has only resulted in a
very faint smear. If anyone knows of a  better method of purifying
bacterial genomic DNA, or if you've had experience working with B.
sterolicum, I'd appreciate some help.

Also... I'm trying to PCR amplify a 1.7 kb segment from the said DNA.
I'm using an Idaho Technologies RapidCycler with 10 ul volume/sample,
using an annealing temp. of 45-50 deg. C. for about a minute. These
temperatures are consistent with my primer's specifications, but I
can't seem to get any product. Any help with this would also be
appreciated.




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