How to store RNA??
gshearer at whale.st.usm.edu
Tue Jun 13 12:21:43 EST 1995
"Aron B. Jaffe" <jaffe at a1.mscf.upenn.edu> wrote:
> cnguan at julian.uwo.ca () wrote:
> > I have generated some RNA by in vitro transcription and would like to
> > store it for later use. I have heard that multiple freeze-thaw cycles
> > breaks down RNA very quickly...is this true and how do I get around
> > it?
> > Thanks!
> > -Chris
: After taking the OD, you can store your RNA under ethanol at a
: concentration you like, and then take out as much as you need
: for each experiment (being sure to mix well by vortexing). You
: have to go through the joy of pelleting and resuspending each
; aliquot, but the RNA is stable this way for a long time.
Be wary about taking out an aliquot of RNA in ethanol. Its an
excellent way to store RNA at -20C but we've found that removing
a certain number of ug of RNA is risky. For example, if you know
from previous A260 reading that you have 100 ug RNA in 100 ul
EtOH soln' then 10 ul should give 10 ug. Don't bet on it! You
may suck up a chunk of RNA ppt and thus have a lot more (or not
suck up some ppt.and have a lot less) RNA that you think.
If its necessary to store RNA and remove accurate quantities
we aliquot RNA in sterile water and store frozen at -70C. Each
tube is thawed only a few times before its empty then we go to
the next tube, etc.
[gshearer at whale.st.usm.edu]
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