RNA resuspension problem
SURESH KUMAR K.G.
skg at biochem.iisc.ernet.in
Tue Jun 19 14:44:46 EST 2001
This could be due to polysaacharides getting into your RNA prep. May be
you are going a bit too close to the interphase while pipetting out the
sup after phenol:chloroform extraction. Try avoiding this..your
RNA will be fine.
Heating for long period is not good.
One more thing you can check...just spin down the gel like pellet..and
check out the sup for RNA.
Hope this helps.
On 19 Jun 2001, Melanie wrote:
> I have in the past had no problems precipitationg and resuspending my RNA in
> DEPC H20. However, the last time I tried to do this using 1/10 volume NaAc
> ph 5.5 and 2 volumes EtOH and freezing at -20 overnight, I come up with a
> clear gel like blob of a pellet that I can't get to redissolve. I have
> heated at 65 for 10 min several times, vortexed, flicked the tubes
> repeatedly, etc. Should I heat the tubes longer? Will that harm the RNA?
> Any other suggestions?
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