Is there a simple test for RNase and DNase free
eooe at hotmail.com
Mon Jan 29 12:00:50 EST 2001
Can you be more specific as to how much buffer and suspect to use? How
about 50 mL buffer with 0.1 gram reagent.?
> DNase can be checked by incubating a plasmid digest of small fragments
> (i.e. 50-500bp) in a 1x LOW RE reaction buffer with the suspect whatever
> for 16 hours at 37C, remembering to do a negative control, as well. Run
> on acrylamide gel and see if there is any change.
> For RNA you can do something similar with a transcript etc.
> For protease, Taq pol is a great one for checking pancreatic DNAse to be
> protease free. It really gets chewed up when visualised on a protein
> gel. Problem is you need 1ug, 250u, per assay. Not a problem for us, a
> manufacturer but I'd guess there must be fluorescent protease substrates
> that could be used in place.
> For increased sensitivity on RNase and DNase use 32P labelled substrates
> and run as above and autoradiograph or better still count released 32P.
> Very sensitive.
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
> Duncan Clark
> GeneSys Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
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