Protocol for conversion of Uridine to Pseudouridine using Sce PUS1 (NEB #M0526)
- Prepare 100 µl reaction as described below:
Components
100 µl Reaction RNA
up to 1 µg
NEBuffer r1.1 (10X)
10 µl
Sce PUS1
1 µl (100 pmoles)
Nuclease-free water
to 100 µl
- Incubate at 30°C for 2 hours
- The reaction may be stopped by the addition of 1 µl of NEB Proteinase K and 15 min incubation at 37°C. Subsequent RNA purification using the appropriate NEB Monarch RNA Cleanup Kit can be used to clean up the RNA for downstream applications.