?best purification of 186bp DNA fragment from agaraose

Ian A. York york at mbcrr.dfci.harvard.edu
Wed Nov 23 16:47:04 EST 1994


>Robert C. Colgrove (robin at alumni.caltech.edu) wrote:
>: I am cloning a small (186bp) PCR product. Usually I clone larger fragments
>: and purify the gel fragments with "geneclean." Their package insert
>: suggests not using it for less than 200bp. In the old days, I used all
>: the standard tricks: Low melt agarose, electroelution, mashing up the 
>: gel slice, etc. I was just wondering what people tihink is the best way
>: to go about it these days. Thanks.

Qiagen's kit for purification from agarose - Qiaex - works down to 
something like 50 bp.  I used it successfully for a 200 bp fragment that 
didn't work well with GeneClean.  However, my impression is that the 
GeneClean is unpredictable at these sizes and it may well work with your 
fragment - it's worth a try.  If not, Qiaex should do the trick.

Ian

-- 
Ian York   (york at mbcrr.harvard.edu)
Dana-Farber Cancer Institute, 44 Binney St., Boston MA 02115
Phone (617)-632-4328     Fax  (617)-632-2627




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