Protocol for conversion of Uridine to Pseudouridine using Sce PUS1 (NEB #M0256)

  1. Prepare 100 µl reaction as described below:

    Components
    100 µl Reaction 
    RNA
    up to 1 µg
    NEBuffer 1.1 (10X)
    10 µl
    Sce PUS1
    1 µl (100 pmoles)
    Nuclease free H2O
    to 100 µl


  2. Incubate at 30°C for 2 hours

  3. 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.

 


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Related Products:
Sce PUS1