RNA extraction

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Fri Jul 14 20:49:49 EST 2000


"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? 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!!
>

If your RNA looks good, not degraded, gives you a good signal,
works for everything you need it for, etc., then stop worrying
about the absorbance ratio and do experiments instead.

Nick


Nick Theodorakis

nicholas_theodorakis at urmc.rochester.edu

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