Radioactive Labeling with T4 PNK or T4 PNK (3´ phosphatase minus)
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1. Set-up the following reaction:| DNA | Up to 50 pmol of 5´ termini |
| T4 PNK Reaction Buffer (10X) |
5 µl |
| ATP (10 mM) |
50 pmol of [γ-32P] ATP |
| T4 PNK | 2 µl (20 units) |
| Nuclease-free Water | up to 50 µl |
2. Incubate at 37°C for 30 minutes.
3. Heat inactivate by incubating at 65°C for 20 minutes.
Note: [33P] ATP may be substituted for [32P] ATP.