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      Radioactive Labeling with T4 PNK (M0201)

      New England Biolabs

      Published: 2022-02-20 DOI: 10.17504/protocols.io.bgtzjwp6
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      Abstract

      Radioactive Labeling with T4 Polynucleotide Kinase.

      Before start

      Steps

      1.

      Set-up the following reaction:

      AB
      ComponentAmount
      DNAUp to 50 pmol of 5´ termini
      T4 PNK Reaction Buffer (10X)5 µl
      ATP (10 mM)50 pmol of [γ-32P] ATP
      T4 PNK2 µl (20 units)
      Nuclease-free Waterup to 50 µl

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

      2.

      Incubate at 37°C for 0h 30m 0s.

      3.

      Heat inactivate by incubating at 65°C for 0h 20m 0s.

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