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

    Protocols.io also provides an interactive version of this protocol where you can discover and share optimizations with the research community. This is available for both the Radioactive Labeling with T4 PNK and the Radioactive Labeling with 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.