One-pot digestion of 5´ triphosphorylated RNA using RppH (NEB #M0356) & XRN-1 (NEB #M0338)
- Set up the reaction as shown below.
Component Volume 5´ triphosphorylated RNA Up to 2 µg NEBuffer 2 (10X) 3 µl RppH (5 U/µl) 1 µl XRN-1 (1 U/µl) 2 µl Nuclease-free water to 30 µl
- Incubate at 37°C for one hour.
- Stop the reaction by adding EDTA to a final concentration of 20mM. Alternatively, you can directly proceed to RNA clean up.