Standard Decapping Protocol (NEB #M0608)
- Set up the following reaction on ice.
COMPONENT 20 µl REACTION FINAL CONC.
mRNA Decapping Enzyme Reaction Buffer (10X) 2 µl 1X Total RNA up to 5 µg mRNA Decapping Enzyme 1 µl 5 units/µl Nuclease-free water to 20 µl
- Gently mix the reaction by pipetting up and down and microfuge briefly.
- Incubate at 37°C for 30 minutes.
- Use a final concentration of 25 mM EDTA to stop the reaction.
- Use a final concentration of 2% SDS to stop the reaction.