A Typical DNase I Reaction Protocol (M0303)
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- Set up the following reaction on ice:
COMPONENTS 100 μl REACTION 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
- Incubate at 37°C for 10 minutes.
- Add 1 µl of 0.5 M EDTA (to a final concentration of 5 mM).
- 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.