Removal of pyrophosphate from 5’ triphosphorylated RNA using RppH (NEB #M0356)
|5’ triphosphorylated RNA||Up to 2 µg|
| NEBuffer 2
|RNA 5’ Pyrophosphohydrolase (RppH)||2 µl|
|Nuclease-free water||to 20 µl|
2. Incubate at 37°C for 30 minutes.
3. Stop the reaction by adding EDTA to a final concentration of 20mM. Alternatively, you can directly proceed to RNA clean up.
Note: The EDTA chelates Mg++ ions so if you intend to follow the RppH reaction with another enzymatic reaction that uses Mg++ without reaction clean up, avoid adding EDTA. We recommend purifying RNA between different enzymatic steps to ensure optimal performance of the enzymes.