sergioal at bbm1.ucm.es
Thu Feb 1 09:44:05 EST 2001
Some time ago we had similar (but not the same) problem with RNase. We get DNA
gel shifting in the RNase treated samples, using the RNase supplied by Roche and
prepared following the Maniatis protocol. The only solution we found was to
dilute the RNase, about 10X less than suggested in the protocol, and the
shifting dissappeared but the RNase still worked. Are you sure you aren't
getting a gel shift?
PS: i remember somebody cited a paper about this RNase DNA-binding.
> We´ve prepared RNase several times (as it is written in MANIATIS) to get it
> Now we´ve tested this RNase on genomic- and Plasmid-DNA (1.5 h, 37
> degree) - a control without RNase-treatment was included.
> The control DNA was o.k., but there was nothing left from the RNase treated
> DNA (in both cases).
> As a next step we´ve tested some old batches of RNase - which have worked
> well all the time before - but the result was the same (control o.k.,
> nothing left from the treated DNA).
> Does anyone have an explanation for this phenomenon?
> Thanx a lot for any answer.
> knobloch at uni-duesseldorf.de
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