DNA recovery from TBE gel

Alex Brands abbrands at artsci.wustl.edu
Wed Jun 24 19:21:28 EST 1998


Hello all,

I'm doing a partial digest of a 10.5kb plasmid, and I need to gel purify
the linear DNA from the uncut.  On a .7% TBE gel, the linear and uncut
bands are resolved nicely.  However, when I ran the digest on a .7% TAE
gel, the bands were not resolved.

Now, I've always heard that you're supposed to do gel purification with
TAE gels, although I don't know what the problem with TBE is.

My questions are;  What is the problem with isolating DNA from TBE gels?
                   Is there some way around the problem?
		   Is there some other solution to my problem?

In case it makes a difference, I need the DNA to do gap repair cloning in
yeast.  The insert I have contains the genomic sequences flanking my gene
with an Ssp1 site in the middle.  Unfortunately all the enzymes that cut
in the right places of the full length genomic clone also cut in the
vector.

Thanks!

Alex




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