ShortCut RNase III Digestion Protocol (NEB #M0245)


  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.