Ribosomal RNA prep?

John McDougall johnmcd at fagmed.uit.no
Fri Sep 23 05:31:20 EST 1994


In article <1994Sep19.151629.27576 at ac.dal.ca>, isbruck at ac.dal.ca (RICHARD
ISBRUCKER) wrote:

> I'm having difficulty probing my northerns since the mRNA that I am
> looking for is so close to that of rRNA, and I am also beginning to
> suspect that the probe is binding non-specifically to rRNA (but am not
> 100% sure about that).  To verify the specificity of the probe, I would
> like to run a control lane of pure ribosomal RNA, but i'm not too sure
> how to isolate it.   Does anyone know of a method for isolating ribosomal
> RNA from tissue or from a total-RNA prep?  I thought about cutting it
> out of a gel, but i'm worried that i'll be getting the mRNA I'm probing
> for in the same piece.  I'm not too sure how well a poly-dT column will
> work since there is the question of non-poly-A-tailed mRNA.  All suggestions
> will be greatly appreciated.
> 
> Thanks,
> --
> Richard Isbrucker                       e-mail: isbruck at ac.dal.ca
> Dalhousie University                    Tel:    (902) 494-2571
> Department of Pharmacology
> Halifax, Nova Scotia, Canada, B3H 4H6


If you want to check out your probe on the rRNA, you will have to isolate
ribosomes first, and then protein extract - presto- rRNA at your finger
tips! :)

-- 

John McDougall       / TEL: 47-776-44479
NFH                  / FAX: 47-776-46020
Univ. of Tromsoe     / E-MAIL: johnmcd at fagmed.uit.no
N-9037 Tromsoe       /
 Norway              /



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