RNase protection assays...

Helmut Dotzlaw dotzlaw at ccu.umanitoba.ca
Sun May 9 12:56:03 EST 1993

In article <1993May9.101615.16630 at news.acns.nwu.edu>,
drmax at casbah.acns.nwu.edu (Marianna Max) wrote:
> In article <5MAY93.11573872 at wums.wustl.edu> wetsel_r at wums.wustl.edu writes:
> >Hi Netters:
> >
> >I noticed that in addition to Ambion, Clonetech is now offering an RNase 
> >protection kit called the Guardian.  Like Ambion's kit, literature from 
> >both company's is quite vague at the step where inactivation of the RNases
> >takes place. (Sounds like proprietary kit information again!)  Having done 
> Ambion's instruction book that comes with the assay contains a complete
> list of all formulations, nothing proprietary at all (I'm refering to the first
> version of the kit not their second version, which I haven't used). The
> inactivation of RNase step in the kit is Prot K and SDS followed by a
> phenol extraction and a percipitation. I've never had problems with the 
> RNA redisolving using this method.

I'm sure David is referring to the RPA II kit, it does not involve Prot K
or phenol extraction.  As to the list of formulations, two of the "magic
juices" are not described - Soln. BX (RNase Digestion Buffer) and Soln. Dx
(RNase Inactivation/Precipitation Mixture (patent pending).

Helmut Dotzlaw            "You can observe a lot by just watchin.."        
Dept. Biochem. Mol.Bio.    -Yogi Berra
University of Manitoba
Winnipeg, Canada

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