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T4 Polynucleotide Kinase catalyzes the transfer of the γ -phosphate of ATP to the 5´ hydroxyl terminus of DNA or RNA. A T4 Polynucleotide Kinase Technical Bulletin is available.
Applications: 5´ end-labeling of DNA or RNA (1). Phosphorylation of synthetic linkers (1).
Source: Purified from E. coli expressing the T4 polynucleotide kinase gene in a λ vector.
Performance and Quality Testing: Ribonuclease, endodeoxyribonuclease, 3´ and 5´ exodeoxyribonuclease, and phosphatase assays; labeling efficiency tested.
Unit Definition: One unit transfers 1 nmol of phosphate of ATP to the 5´ hydroxyl termini of micrococcal nuclease-treated DNA in 30 min. at 37°C.
Unit Reaction Conditions: 70 mM Tris-HCl (pH 7.6), 0.1 M KCl, 10 mM MgCl2 , 1 mM 2-mercaptoethanol, 0.3 mM [γ-32P]ATP, 0.38 mg/ml micrococcal nuclease-treated DNA, and enzyme in 150 µl for 30 min. at 37°C.
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