RNase removal by phenol-chloroform ?

Edmundo Castro ecastro at physci.ucla.edu
Fri Dec 6 22:19:20 EST 1996


nierlich at ucla.edu wrote:
> 
> In <57utfc$69g at mserv1.dl.ac.uk>, deepak at bgumail.bgu.ac.il (Deepak Khandka) writes:
> >Is  it possible to remove RNase contamination from a Tris-containing buffer
> >by extracting with phenol-chloroform without purturbating the pH of the
> >buffer ?

> 
> One way to make your buffer RNase-free is to treat with diethylpyrocarbonate, as described in Maniatis.  Add (I think)
> 0.1 % DEPC to the buffer, mix, invert the bottle with a slightly loose cap so the neck is wetted, let stand for a few
> hours--overnight, autoclave (to drive off residual DEPC/alcohol/CO2), and store until needed.
> 
> Don Nierlich
> >

Tris solutions are NOT to be treated with DEPC.  See Maniatis

E. Castro-Vargas



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