RNA extraction

Jaco Hendriks jaco.hendriks at hetnet.nl
Sun Jul 16 03:00:33 EST 2000


Hi Tina
try to wash a second time with large amounts of ethanol. It seems to me the
pellet is salty.
Good luck
Brenda

"Tina" <anfortin at webnet.qc.ca> wrote
news:vkPb5.1011$WM6.75101008 at news1.mtl.metronet.ca...
> 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? I have already read that to calulate the ratio it is
> better to take ODs of RNA in TE buffer but other people have already got
> good ratios with RNA in water.
>
> Any suggestion?
> Thanks a lot!!
>
>







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