Can anyone help with dephosphorylation of RNA using Rpph?

13 July 2014 1 10K Report

Hi,

I am looking for a working protocol to dephosphorylate the 5'PPP into a 5'P using NEB Rpph (https://www.neb.com/products/m0356-rna-5-pyrophosphohydrolase-rpph). Do you denature RNA before dephosphorylation? Do you purify this using the RNA clean and concentrate kit?

I am using 15ul(RNA+dH2o) , 4ul of NEB buffer 2 and 2ul of Rpph for a 20ul reaction. I purify it using LiCl precipitation.

Has anyone also used T4 RNA ligase 1? It would be helpful.

Ishita Jain

Did this protocol work for you?

https://benchling.com/protocols/6fCizH/dephosphorylation-of-5ends-of-rna

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