T4 Polynucleotide Kinase (PNK)

Polynucleotide Kinase (PNK) exhibits 5’ polynucleotide kinase and 3’ phosphatase activity. It catalyzes 5’-phosphorylation of DNA and oligonucleotides via transferring the terminal phosphate of ATP to a 5’ hydroxyl group of a nucleic acid.

Source

E. coli strain expressing a cloned pseT gene of bacteriophage T4.

Includes                      

• T4 Polynucleotide Kinase (PNK)
• 10X PNK reaction buffer

10X PNK Reaction Buffer

500 mM Tris-HCl, 100 mM MgCl₂, 50 mM dithiothreitol, pH 7.5 @ 25°C.

Storage Buffer

50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC.

Unit Definition:

One unit of T4 Polynucleotide Kinase converts 1nmol of ³²P from [γ-³²P]-ATP into an acid-insoluble form in 30 minutes at 37°C under standard assay conditions.

Molecular Weight

Theoretical: 34.6 kDa

Storage temperature

-20°C.

Inactivation

Inactivates by heating at 70°C for 15 min.

Inhibition

Metal chelators, phosphate and ammonium ions, KCl and NaCl at a concentration higher than 50 mM can inhibit PNK

Quality Control Assays

T4 PNK is free from detectable nuclease and RNase contamination.