Page purification of large oligos: resolving 50mer from 49mer?
Gys de Jongh
GysdeJongh at planet.nl
Tue Oct 30 15:31:25 EST 2001
"Lucy McGillicuddy" <lucy_mcgillicuddy at hotmail.com> wrote in message
news:d6bb0d4d.0110291304.a039a69 at posting.google.com...
> I'm hoping someone can give me some advice about purifying large
> oligos. I'm PAGE purifying 50mers, and having trouble eliminating n-1
> products. I'm loading 2-4 OD units on 12% acrylamide gels. Rather
> than distinct 50mer and 49mer bands, I get a large, rather long blob.
> I thought maybe we're loading too much DNA, however, my supervisor
> believes that if this were the case, we would have two distinct blobs
> rather than one. We've tried 14% gels, and this doesn't make much
> difference in resolution. I looked into precast gradient gels, but
> these don't come in sizes large enough to run 50mers. I'd really
> appreciate any ideas on how to separate 50mers from 49mers.
we used 12 % gels to resolve 10 - 100 nucs. I think the resolution is too small
49 from 50. Maybe if you let it run for 20cm (6 h). Do you have enough ureum to
make it all ss ?
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