Alkaline gels-Better method?
David T Wong
wong at cardinal1.Stanford.EDU
Thu Feb 29 19:58:05 EST 1996
I'm trying to separate PCR products on 4% Alkaline agarose gels with NaOH
and EDTA. If I run them overnight the bands are diffuse. If I run them
over 5 hours (400mA and about 90V in BioRad wide minisub cells) with lots
of buffer cooling, I get uneven running. The top of the gel with cool
buffer runs faster than the bottom on the tray. I have to take pictures
at a 45 degree angle. Urea gels don't work for me.
Does anybody have a better way of running alkaline gels at reasonable
field strengths. Is there another high pH buffer system that is
compatable with agarose or acrylamide (acrylamide hydrolyzes in the
NaOH/EDTA buffer). Is there a more stable gel? I read about AAEE
acrylamide but where can I buy it? Any suggestions would be appreciated.
Division of Gastroenterology
wong at leland.stanford.edu
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