Page purification of large oligos: resolving 50mer from 49mer?

Lucy McGillicuddy lucy_mcgillicuddy at hotmail.com
Mon Oct 29 16:04:51 EST 2001


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.

Thanks,
Lucy




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