Subcloning dirty PCR products
Ian A. York
york at mbcrr.dfci.harvard.edu
Fri Aug 26 10:04:46 EST 1994
In article <33iv77$la3 at u.cc.utah.edu> Rafael at genetics.med.utah.edu (Rafa Maldonado) writes:
>In article <1994Aug24.231015.1 at mcclb0>
>rosas at mcclb0.med.nyu.edu writes:
>> 2. Is it possible to get ride of such by-products without repeating
>> the PCR reaction (I'm short of primers), say by cutting my PCR product
>> out of a LM agarose gel???
>Yes, you can try some gel-extraction system, using high resolution
>agarose and a very good system of recovery from the gel (as quiagen
>beads). But I prefer centricon 100. If you need special protocols,
You can also try purifying with Geneclean directly from the PCR mix,
depending on your product and the contaminants. Geneclean is inefficient
at purifying products less than about 250 bp, so it will not pull out the
primers or short contaminants which might be inhibiting your subsequent
reactions. Qiagex is more efficent for short products (literature claims
down to 50 bp or less, I think) which might be a disadvantange.
The last package of Geneclean II I saw new had an insert in it
discussing purification from PCR.
Hope this helkps.
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