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T4 Polynucleotide Kinase

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • T4 Polynucleotide Kinase catalyzes the transfer of the ?-phosphate from ATP to the 5´-terminus of polynucleotides or to mononucleotides bearing a 5´-hydroxyl group. The enzyme may be used to phosphorylate RNA, DNA and synthetic oligonucleotides prior to subsequent manipulations such as ligation and cloning. T4 PNK is provided with 10X Reaction Buffer: 700mM Tris-HCl (pH 7.6 at 25°C), 100mM MgCl2, 50mM DTT.

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    Storage Buffer: 20mM Tris-HCl (pH 7.5), 25mM KCl, 2mM DTT, 0.1mM EDTA, 0.1uM ATP and 50% (v/v) glycerol.

    Source: Recombinant E. coli strain.

    QC Tests: Activity, DNase, RNase, endonuclease, end-labeling, blue/white cloning assay.

    Unit Definition: One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of phosphate to the 5´-OH end of a polynucleotide from [?-32P]ATP in 30 minutes at 37°C. The reaction conditions are: 40mM Tris-HCl (pH 7.5 at 25°C), 10mM MgCl2, 5mM DTT, 0.1mM [?-32P] ATP and 0.5mM 5´-OH polynucleotide end concentration.

  • Protocols

    Complete Protocol

    Download PDF

    T4 Polynucleotide Kinase Technical Bulletin

    PDF (144 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources