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.