Why can't I get agarose embedded HMW DNA to digest ?

Howard T. Petrie h-petrie at ski.mskcc.org
Fri Mar 17 11:56:02 EST 1995


In my experience, which is now fairly extensive, some enzymes just won't
cut in solidified agarose.  Most of these (all in my experience) will cut,
however, if you melt the agarose first, cool to 37C, and then add enzyme. 
Also, minimization of the [agarose] will help facilitate digestion.

In article <3kbn92$mfk at is.bbsrc.ac.uk>, cavell at BBSRC.AC.UK) (A Cavell) wrote:

> I am trying to do some long range physical mapping of the Brassica genome.
> I obtain high moleclar weight DNA by embedding protoplasts in low melting
> point agarose... I get little or no digestion.



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