RNA folklore?
Cornelius Krasel
krasel at alf.biochem.mpg.de
Wed Dec 14 17:51:51 EST 1994
hrasal at marc.cri.nz wrote:
> There are two things that I'm curious about. When I first started
> working with RNA, I was told that autoclaving inactivated RNAses, but the
> RNAse protein will (after a certain period, 24 hours or so), reform into the
> active enzyme. I had assumed that this was stated in Stryers Biochemistry.
> I was wrong. Actually, it was denaturation of ribonuclease in 6M guanidine
> HCl followed by removal of the guanidine by dialysis. So, is reactivation
> of RNAse activity after autoclaving an urban legend, or is it real?
I don't know about autoclaving but to get DNase-free RNase you make a
solution of cheap RNase A (which contains DNase) and boil it for a while.
When cooling down, the RNase will renature, the DNase will not. For
details see Maniatis.
--Cornelius.
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/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */
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