ShortCut RNase III Digestion Protocol (NEB #M0245)

Protocol

1. Set up a 100 μl reaction as follows: 
   

   Nuclease-free Water	70 µl – x µl
   10X ShortCut Reaction Buffer	10 µl
   dsRNA	x µl (10 µg)
   10X MnCl2	10 µl
   ShortCut RNase III	10 µl
   Total Volume	100 µl

2. Mix and incubate for 20 minutes at 37°C.
3. Add 10 µl 10X EDTA to stop the reaction. Do not heat inactivate. Heat inactivation reduces the yield of siRNA.