Solubilizing RNA in water

Matthew L. Beyers mbeyers at achilles.net
Sat Jun 6 08:55:29 EST 1998




> > Mary_Wood at BAYLOR.EDU wrote:
> > >
> > > I've been using a product called Tri-Reagent to isolate RNA from tobacco plants.
> > > I get large amounts of RNA but often the pellet is insoluble in water after the
> > > 75% ethanol wash, even when I don't dry the pellet at all.  I need to be able to
> > > go into an RT-PCR with my sample.  Does anyone have a solution?
>
> Hi!
>
> We routinely dissolve our plant RNA at 65 deg. for half an hour.
> After that centrifuge and transfer supernatant to a fresh tube.
>
> Ricky

Just thought I'd add my two cents.  I do trout RNA isolations routinely and I use a
GITC/phenol/chloroform method which gives me terrific results and large yields (always a
ratio of ~2.0 for A260/A280) but I have this problem of dissolving as well and I
sometimes put one sample in 65 C for ~10 min. while I work on dissolving another and I
switch them back and forth.  I never leave one sample in too long (could lead to
degredation) and I always put it on ice when it's fully dissolved.  Works every time for
me.

Good luck!

Matt






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