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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