T4 Polynucleotide Kinase

T4 Polynucleotide Kinase catalyzes the transfer of the γ-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, 0.1μM 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 [γ-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 [γ-³²P] ATP and 0.5mM 5´-OH polynucleotide end concentration.