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  • Radioactive Labeling with T4 PNK or T4 PNK 3' phosphatase minus

    Set-up the following reaction:

    DNA Up to 50pmol of 5' termini
    10X T4PNK Reaction Buffer 5 µl
    ATP 5 μl of 50 pmol of [γ-32P] ATP µl
    T4PNK 2 µl (20 units)
    Water up to 50 µl

    Incubate at 37°C for 30 minutes.
    Heat inactivate by incubating at 65°C for 20 minutes.

    Note: [33P] ATP may be substituted for [32P] ATP.