Recombinant T4 Polynucleotide Kinase (T4 PNK).
•Applications include end-labeling and insert preparation for cloning.
T4 Polynucleotide Kinase catalyzes the transfer and exchange of gamma-phosphate of ATP to the 5´-hydroxyl terminus of polynucleotides (double-and single-stranded DNA and RNA) and nucleoside 3´-monophosphates.
Purified from a strain of E. coli that carries a T4 Polynucleotide Kinase expression plasmid.
• End-labeling DNA for probes and DNA sequencing (1)
• Addition of 5´-phosphates to oligonucleotides to allow subsequent ligation (2)
Heat inactivation: 70°C for 15 minutes.
Enzyme and buffer should be stored at -20°C. Enzyme is stable for 12 months if handled properly.
• Ammonium ions are potent inhibitors of T4 Polynucleotide Kinase. DNA should not be precipitated from or dissolved in buffers containing ammonium salts.
• Protruding 5’-overhangs are more efficiently phosphorylated than blunt- or recessed 5’ overhangs.