How to remove diphosphate from in vitro transcribed RNA? - (Feb/02/2010 )

I want to prepare in vitro transcribed capped RNA and I know that the capping eficiency is around 80%. So my goal is to remove RNAs without cap from reaction mixture.

There is a Terminator™ 5´-Phosphate-Dependent Exonuclease from Epicentre Biotechnologies that digests RNA that has a 5´-monophosphate end. It does not digest RNA that has a 5´-triphosphate and this is the problem because in vitro transcribed RNAs have 5´-triphosphate.

So is there any possibility to remove diphosphate from 5' end of uncapped RNAs? If it is possible I could use this enzyme


best regards

blaszcz

ok i know how to:)

the same company has RNA 5´ Polyphosphatase. The enzyme sequentially removes the γ and β phosphates from 5´-triphosphorylated RNA


cheers