Standard Decapping Protocol (NEB #M0608)

  1. Set up the following reaction on ice.

    COMPONENT 20 µl REACTION FINAL CONC.
    OR AMOUNT
    mRNA Decapping Enzyme Reaction Buffer (10X) 2 µl (1X) 1X
    Total RNA up to 5 µg  
    mRNA Decapping Enzyme 1 µl 10 units/µl
    Nuclease-free water to 20 µl  
  2. Gently mix the reaction by pipetting up and down and microfuge briefly.

  3. Incubate at 37°C for 30 minutes.

Note: The enzyme can be inactivated with EDTA or SDS.

  • Use a final concentration of 25 mM EDTA to stop the reaction.

  • Use a final concentration of 2% SDS to stop the reaction.
The reaction volume can be scaled as needed. The incubation time can be increased to 1 hour if necessary.