lysis buffer preserving protein-RNA interactions

Pow Joshi via methods%40net.bio.net (by pow.joshi from gmail.com)
Fri Mar 2 10:47:46 EST 2007


On 2 Mar 2007 04:10:56 -0800, Lechu <lech_kaczmarczyk from yahoo.com> wrote:
> Hi all,
>
> Could anyone recommend me some lysis buffer for brain tissue,
> preserving RNA-PROTEIN interactions. I plan to do mRNA 'pull-downs'
> with rec HisTagged RNA binding protein. Want to use both, isolated
> mRNA and crude cell lysate.
> HEPES-based ELB buffer will do? Any other suggestions?

ah, I did some protein pull down assays with the brain tissue ....
works well with the Hepes +NP-40/TritonX +protease inhibitors and
EDTA.... I suppose you would have to add some DTT/DEPC and Rnase
inhibitors, to preserve your RNA ......

best
pow

> Thanks in advance,
>
> Lech
>
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