RNasing minilysates

William R. Morgan wmorgan at ACS.WOOSTER.EDU
Wed Jul 28 15:30:39 EST 1993


In an apparently truncated letter, beatriz at pine.circa.ufl.edu writes:
>When I do my minilysates I usually digest with about 2 ul of 20 mg/ml of RNase
>H after the enzyme restriction digestion. The problem I've had is that if I'm
>not careful and pipet more than 4 ul or so, my samples don't even get into 
>the gel (0.8% agarose-TAE). I get these huge aggregates that stay in the
>wells. It is so annoying lately I just run my gel and then RNase the whole
>gel, but this takes much longer and the bands from the inserts are usually
>somewhat diffuse. Has anyone see
>

oo

I usually add RNAse to a concentration of ~20 micrograms/ ml before I set
up my restriction digests, so that the RNA is degraded during the ensuing
restriction digest.  It works fine.

Bill Morgan


William R. Morgan
The College of Wooster
Department of Biology
931 College St.
Wooster, OH  44691
Phone:  216-263-2026
FAX:    216-263-2378
E-mail: wmorgan at acs.wooster.edu




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