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siRNA extraction from plant material - (May/12/2006 )

Does smallRNAs were precipitate by Ethanol or Isopropanol? Which extraction protocol works good to isolate small RNAs?

-poplar-

hi
tri reagent or trizol work good for siRNA extraction. EtOH and IprOH precipitaes both ethanol if added in enough amount. (1:1 vol per isoprop, 1:2,5 per ethanol (2,5 volumes of ethanol of course))
you may add glycogen or tRNA as carriers.

-fred_33-

Is it necessary to keep the sample at -20 C (as for Etof precipitation) when using Isopropanol to precepitate small RNA? Few publications recommend to keep the sample at -20 C for 30 min and others to keep at room temp. for the same span of time to precipitate the RNA. Which is better for iso propanol precepitation?
Thanks,
Nazmul

-Nazmul-

i incubate them RT when using IprOH. But 30' at -80 when using EtOH.

-fred_33-

Can you please explain why do you keep the samples at RT while using IsoOH?
Thank.

-Nazmul-

it's related to precipitation capability of the two molecules.
The more C in the molecule, the more precipitation efficiency occurs.

you may have a look at these threads :

dielectricity and ethanol precipiation

iproh etoh and temperature

-fred_33-