IUBio

RNA

yoel and adaia shiboleth shibolet at mishkei.org.il
Sun Mar 28 17:17:40 EST 1999



Olivier Lefebvre wrote:

> Hi
>
> I am looking for a simple protocol to extract only cytoplasmic RNA from
> frozen tissu!
>
> Thanks in advance
>
> Olivier Lefebvre
> de France Tel: 03 88 27 77 27
>               Fax: 03 88 26 35 38
> From abroad: Tel: 33 3 27 77 27
>                     Fax 33 3 88 26 35 38
> Adresse: U.381 INSERM
> 3 Avenue Molière
> 67200 Strasbourg
> France
> E-mail: Olivier.Lefebvre at inserm.u-strasbg.fr

Hi
Red-book or maniatis contain a method for purifieing nuclei. The sup
containes the cytoplasmic RNA.
The method uses Tris, MgCl2, NP40. You add DTT, spermidine and RNAsine (a
lot, very expensive...).
centifugation is 500g 5' twice.
I don't remember the concentrations by heart though I made the solution
today.
High grade nuc/cyto RNA is achieved from many different tissues and cell
lines.
Yoel




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