Redissolving RNA Pellet

clarosa clarosa at biocomp.unl.edu
Thu Sep 14 12:49:41 EST 2000



Gina Beck wrote:

> I am working with total RNA from Chondrocyte cells.  I am isolating the
> RNA using TRizol Reagent and I am having a problem with the last step of
> redissolving the RNA pellet.  These cells have been put in 1.2% alginate
> and cultured for 5 days and then extracted out of the alginate using 55
> uM of Na Citrate solution.  Then I add Trizol and continue with the
> isolation.  Do you think there may be excess of proteoglycan material
> that is making the pellet hard to redissolve?  any suggestions?
> Heather

Be careful not to dry the pellet completely as this makes the rna much
harder to dissolve.  If there is a little residual 70 percent ethanol I
dont think it hurts. In my use of trizol with Dicty or plant materials I
always observed a residual pellet that never dissolves.  After a few
qualms, I learned to ignore the pellet and so far northerns have worked
reasonably well.   You may have to play with the rna (mash it , vortex) and
dissolve it over a long period with some heating.  I turned to dissolving
in formamide with eliminates worries about residual RNASES.







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