Ribosomal RNA prep?

Tracy Aquilla aquilla at salus.med.uvm.edu
Mon Sep 19 16:22:53 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

I haven't tried this, but maybe you could use something like DynaBeads to
bind up the mRNA, remove the beads with the mRNA using the magnet, then
precipitate the supernatant containing the rRNA. A couple of rounds of this
procedure should remove the mRNA from your prep, yielding relatively pure
rRNA, shouldn't it?

Tracy Aquilla, Ph.D.
Molecular Physiology and Biophysics
University of Vermont
aquilla at salus.med.uvm.edu



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