Q: Best method for precipitation of small RNA's ( 10-20nt ) ?
Claus Stefan Voertler
voertler at exmed1.gwdg.de
Thu Jun 13 15:49:17 EST 1996
you are really trying to push to the limit here...:-)
Our experience in the lab is that anything bigger than 15-20nt will
precipitate under the normal conditions (1/10vol 3M NaAc, 2.5vol
EtOH) Increasing the conc of NaAc may helped in some cases (to
1/3vol). 2-3M NH4Ac works equally well. A coprecipitant is optional
(glycogen or bulk tRNA) but you have anyway a lot of other nucleic
acids arround. So there are good chances of precipitating a 10 to
15mer with these conditions - might just not be quantitative.
In any case: Why not loading the material directly onto the dPAGE-gel?
If you need to precipitate in order to concentrate the sample, you
could also use ultrafiltration. This works also fine to remove
salts/buffer-components interfering with the electrophoresis...
Claus Stefan Vörtler MPI für experimentelle Medizin
> Hi there !
> I'm doing transcription reactions with nuclear extracts. I'm searching for
> very short transcripts. So what is the best method to ensure precipitation of
> this trasnscripts and analyse them by PAGE after that ?
> Gerd Emde
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