T4 Polynucleotide Kinase

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.

Source:
Purified from a strain of E. coli that carries a T4 Polynucleotide Kinase expression plasmid.

Applications:

• 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.

Storage:
Enzyme and buffer should be stored at -20°C. Enzyme is stable for 12 months if handled properly.

Additional Information:

• 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.

References:

1. Sambrook, J. and Russell, W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), 9.55-9.56, 9.68-9.69.
2. Sambrook, J. and Russell, W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), 13.16-13.17.