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Star Republic: Guide for Biologists

RNA purification --- DNase I digestion

1. Make up the following mix:

RNA (contaminated with DNA) 1-2 mg
10X DNase buffer
(500 mM Tris-HCl, pH 8.0, 50 mM MgCl2 10 mM DTT)
2 ml
RNase inhibitor 1 ml (10 units)
DNase I 0.5 Kunitz units
RNase-free ddH2O to 20 ml

2. Incubate at 37 C for 30 minutes.

3. Stop the reaction by adding 1 ml of 0.5 M EDTA.

4. Heat inactivate the DNase I at 65 C for 5 minutes.