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

cavell at BBSRC.AC.UK cavell at BBSRC.AC.UK
Fri Mar 17 05:11:46 EST 1995

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. These are then lysed and washed extensively using PMSF.

I follow standard protocols for digestion with rare cutting REN's but 

there seems to be a problem with diffusion of enzyme into blocks as I 

get little or no digestion.

Any Ideas ??

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