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