RNase removal by phenol-chloroform ?

Bo Pontoppidan bo.pontoppidan at cellfo.slu.se
Tue Dec 10 15:14:05 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
> >
You should not use DEPC with TRIS!

Bo

--
Bo Pontoppidan                                  Phone: (+46) 018 67 33
24
Uppsala Genetic Center                            Fax: (+46) 018 67 33
89
Dept. of Cell Research                     Home Phone: (+46) 018 69 28
38
Swedish University of Agricultural Sciences, Box 7055,  
S-750 07 Uppsala, Sweden             E-mail:
bo.pontoppidan at cellfo.slu.se



More information about the Methods mailing list