PCR Clean up, 150bp frag???

Tracy Aquilla aquilla at salus.med.uvm.edu
Thu Sep 22 15:31:45 EST 1994


In Article <35s9iiINN6ql at early-bird.think.com>, york at mbcrr.dfci.harvard.edu
(Ian A. York) wrote:
>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.
>

You can also use the MerMaid system, which is made by Bio-101 (GeneClean
guys). MerMaid is designed to purify oligomers, and I have had good success
with it.

Tracy Aquilla, Ph.D.
Molecular Physiology and Biophysics
University of Vermont
aquilla at salus.med.uvm.edu



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