nicholas_theodorakis at urmc.rochester.edu
Tue Nov 16 12:59:04 EST 1999
In article <80s2hv$kar$1 at news.fas.harvard.edu>, Glover Martin
<gwmartin at fas.harvard.edu> wrote:
> Mano <mdep at musica.mcgill.ca> wrote:
> > YES, I am trying to localize a specific message. However, i am
> > wondering what to expect to see by ethidium bromide staining. In
> > fractions would I see the most intense 28 and 18 S rRNAs?
> At the top of the gradient (mostly RNP) you'll see small (tRNA?)
> Then 18S rRNA, coinciding with the 40S subunit. both 18S and 28S
> present in subsequent fractions (even the one containing 60S, in my
> experience). You may be able to see several other bands, by
> EtBr and Methylene blue as well.
I usually add a few micrograms of tRNA as carrier to ppt, so I get tRNA
in every fraction ;-) (Actually, I guess there should be a molar ratio
of 1-2 tRNAs per translating ribosome in the polysome fractions, but of
course by weight % they would be negligible.)
But you're right about the separation of 60S from 80S; it's easy to see
the separation by absorbance peak, but in the gradient fraction I've
never had just 28S rRNA either.
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