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...
> Hello,
>
> 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.
>


HI,
we used 12 % gels to resolve 10 - 100 nucs. I think the resolution is too small
for separation
49 from 50.  Maybe if you let it run for 20cm (6 h). Do you have enough ureum to
make it all ss ?
hth
--
Gys






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