PCR Clean up, 150bp frag???
Ian A. York
york at mbcrr.dfci.harvard.edu
Thu Sep 22 10:58:10 EST 1994
In article <CwHn85.CLE at news.unr.edu> jb at scs.unr.edu (Jon Burrows) writes:
>Can anyone advise me on the most efficient way to clean up a 150bp PCR
>product from multiple rxns..I'm trying to get 50micrograms. Typical resin
>systems don't work well <3% recovery and I'm loathe to cut bands or DEAE
>from a prep gel, although this may be the way to go.
Although the GeneClean resin doesn't work very well below about 200 -
300 base pairs (apparently it will sometimes, unpredictably) the Qiagen product
Qiaex will purify down to about 50 bp or so. I used this directly on my
250 bp product and got nice clean ligatable product; I also ran some on agarose
and purified from a band, which also worked. You also have the option
of purifying from acrylamide gels, which involves just an overnight incubation.
I suppose you could also run it on agarose, cut out a band, and digest
away the agarose with beta-agarase; haven't tried this but it should work.
But the simplest, as far as I'm concerned, is Qiaex.
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