RNA extraction

Chris LaRosa clarosa at biocomp.unl.edu
Thu Jul 20 16:22:51 EST 2000



"David F. Spencer" wrote:
> 
> In article <vkPb5.1011$WM6.75101008 at news1.mtl.metronet.ca>, Tina
> <anfortin at webnet.qc.ca> wrote:
> 
> > Hi everybody!
> > I have isolated total RNA from non adherent cells with the TRIZOL reagent.
> > At the end of the procedure, I solubilized the pellet with DEPC-treated
> > water. After this, I measured OD 260 and 280 and calculated the ratio
> > 260/280. This ratio should be 1.6 to 2.0. Sometimes I have got around 1.6
> > but generally 1.4 to 1.5  ... it is too low but the RNA is OK on agarose gel
> > (no apparent degradation). Which step of the Trizol method can I improve to
> > increase this ratio?
> 

It is possible to obtain a ratio of at least 1.9.  As I did a few hours
ago.  For one , do not be greedy.  When removing the aqueous phase from
the phenol phase , be careful not to try to get all of it.  Also you can
increase the trizol to sample ratio and improve the ratio.






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