How to detect dsRNA? - (Feb/09/2007 )
Let say two complementary 500nt ssRNAs are transcriped simutaniously in cells. How to detect the dsRNA? How to distinguish dsRNA from ssRNA? Does trizol work for dsRNA extraction? Is there any RNase able to digest ssRNA specifically but not dsRNA? Can Northern distinguish the two species? Thanks
So far I know Rnase A only degrades single stranded RNA so you can use Rnase A,
I think trizol may work but i'm not sure,
You might try...
Digestion: 15 min at room temperature using 5 ug/mL RNase A
in 2x SSC: ssRNA is digested, dsRNA is not digested
in 0.1x SSC: dsRNA is digested
20x SSC: 3 M sodium chloride + 0.3 M sodium citrate in water, pH 7.0
Literature: Hull (1985) Purification, biophysical and biochemical characterization of viruses with especial reference to plant viruses. In: Mahy (ed.) Virology - a practical approach. IRL Press Ltd. Oxford, UK, p. 1 - 24
It worked for me.
hi can u tell me how did u extracted dsRNA, basically the methodology which u might have used???