PAGE questions: long run times and streaking
Michael Witty
mw132 at mole.bio.cam.ac.uk
Tue Jan 29 12:07:31 EST 2002
On 28 Jan 2002, Deva wrote:
> 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?
Sounds normal to me. Why not run the gel overnight (if you apparatus does
not leak!).
> 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?
Havn't heard of this. Can you give us a citation? Regards, Mike.
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