hi, i just want to know how to pool RNA? and what should i do before i pool them? i want to pool RNA from several aliquots in order to get a sufficient yield. Will the pooling will cause some decrease of purity? thanks
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Posted 21 July 2009 - 03:25 AM
wntiong, on Jul 21 2009, 04:04 AM, said:
hi, i just want to know how to pool RNA? and what should i do before i pool them? i want to pool RNA from several aliquots in order to get a sufficient yield. Will the pooling will cause some decrease of purity? thanks
You already have a finite amount of RNA so pooling your aliquots will not increase your yield. Do you mean you want a more concentrated stock? If so, pool your aliquots and then do an ethanol precipitation (add 0.1 volume of 3M sodium acetate (pH 5.2) and 2.5 volumes of ice-cold ethanol, incubate at -20 for at least 30 mins) and resuspend the pellet in a smaller volume.
The pooling will only cause a decrease in purity if one of your aliquots is very much less pure than the others - check the 260:280 ratio first,
P
