RNases A in sol. 1
meador at agouron.com
Sat Nov 26 21:54:38 EST 1994
In article <Pine.3.89.9411231655.A18965-0100000 at gadwall.cs.uoguelph.ca>,
lgood at uoguelph.ca (Liam E Good) wrote:
> Several alkaline lysis plasmid prep. protocols have been posted where the
> first solution (sol. 1, PS1, etc., which is used to resuspend the
> pelleted cells) contains RNase A. Does anyone know the first reference
> for this use of RNases A. Also, does the procedure work because the
> enzyme enters the cells and degrades the RNA before the cells are lysed
> with NaOH/SDS? I can't imagine that the RNase keeps working after the
> NaOH/SDS is added.
> Thanks in advance. Please reply to me, because my subscription has been
> getting interrupted.
I am pretty sure that the rnase renatures upon KAc addition after
denaturing in the NaOH/SDS solution. It's a tough enzyme. It's also tough
to get rid of. I don't believe there is a reference.
Hope this helps,
meador at cts.com
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