RNase without RNase
sergioal at *NOSPAM*solea.quim.ucm.es
Thu Feb 17 13:46:52 EST 2000
What kind of problem you've had?
We have been using RNAase A for years without any problem, but recently the digestions
with RNAase produce a DNA shift in the agarose gels. We've tried RNAase from Roche and
Pharmacia without any difference. Has anybody had the same problem?. The only solution we
found is to purify the DNA after digestion, using phenol.
We prepare the RNAase by the Maniatis method: 10 mg/ml in 10 mM Tris pH 7.5 15 mM NaCl.
Boiling 15 minutes and cooling down slowly.
Thanks in advance
Sorry for my bad english
Jim Kami wrote:
> We've had a problem in our lab recently with our genomic DNA
> isolations. Turns out, the cause is the RNase A we've been using.
> Changing suppliers and prep. procedures hasn't helped. I was wondering
> if anyone knows of a way to eliminate RNA from genomic DNA preps without
> using RNase ?
> Thanks in advance.
> Jim Kami
> Dept. of Agronomy
> University of California @ Davis
> Davis, CA 95616
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