PAGE questions: long run times and streaking

Deva dedelman at egeabiosciences.com
Mon Jan 28 20:02:36 EST 2002


I've been using 12% acrylamide gels to PAGE purify 50mer oligos.  I
recently switched to 15%, and I'm getting better resolution, but my
run times are much longer.  I have to wait 7-8 hours as opposed to 4-5
for the oligos to migrate long enough for good separation.  Is this
normal?

Also, I have some streaking above my 50mers that I was able to cut
down on by reducing the voltage.  I read one protocol that suggested
overlaying the sample with methanol, because high molecular weight
nucleic acids tend to get trapped at the surface of the gel well.  Has
anyone tried this methanol overlay technique?

Thanks in advance for any advice!




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