Re: DNase and 2D

dorian immler (dorianimmler@yahoo.com)
Tue, 16 Mar 1999 02:30:27 -0800 (PST)

Removing DNA is not necessary in most cases, especially as you work with
small amounts of material as on analytical gels (i.e. 50 - 250 =B5g).
However, an easy way to get rid of DNA is to increase Ampholyt
concentration to >=3D 0.8% (v/v) in the sample buffer and to precipitate
DNA by doing so. it will form a translucent pellet after
ultracentrifugation.=20

reference:

rabilloud et al., J. Chromatogr. 1986, 351, 77-89

good luck,

dorian

Date:
Mon, 15 Mar 1999 14:48:19 -0300 (DST)
From:
jaime paba martinez <bucaros@guarany.cpd.unb.br>Add to Address
Book
Subject:
DNase and 2D=20
To:
Recipients of ABRF List <abrf@aecom.yu.edu>

I have been reading protocols for protein solubilization in 2D, but=20
they do not talk too much about how to eliminate the contaminant DNA in
the=20
sample. Most of them solubilize the cell extract directly in the=20
rehydration buffer without any other treatment. So, Is it normally=20
enough? does the cell extract becomes just a little=20
viscous? If not, I would appreciate if someone gives me a good
reference=20
or protocol to use DNase and RNase in the procedure.

Thanks in advance,

Jaime Paba
University of Brasilia
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