A Typical DNase I Reaction Protocol (M0303)

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  1. Set up the following reaction on ice:
     RNA ~ 10 μg RNA
     DNase I Reaction Buffer (10X) 10 μl (1X)
     DNAse I (RNase-free)  1 μl (2 units)
     Nuclease-free H2O  to 100 μl
  2. Incubate at 37°C for 10 minutes.
  3. Add 1 µl of 0.5 M EDTA (to a final concentration of 5 mM).
  4. Heat inactivate at 75°C for 10 minutes.

Note: When using RNA in downstream applications, column purification with NEB's Monarch RNA Cleanup Kits (NEB #T2030, #T2040, #T2050), or cleanup with phenol/chloroform extraction is recommended instead of heat inactivation.