T4 Polynucleotide Kinase (10000 U)

A recombinant enzyme from QIAGEN, T4 Polynucleotide Kinase (PNK) catalyzes the transfer and exchange of the terminal gamma-position phosphate of ATP to the 5′-hydroxyl terminus of double- and single-stranded DNA, RNA, and nucleoside 3′-monophosphate molecules. The enzyme also exhibits 3′-phosphatase and 2′,3′-cyclic phosphodiesterase activities and is widely used for 5′-end phosphorylation in labeling and ligation experiments, as well as for studies of nucleic acid repair.

Purified from an E. coli strain expressing the recombinant T4 PNK gene, the enzyme has a molecular weight of 34.6 kDa, a specific activity of 133,333 U/mg, and SDS-PAGE purity greater than 99%. Quality control testing reports single-stranded exonuclease activity of less than 5.0% release, double-stranded exonuclease activity of less than 1.0% release, no detectable double-stranded endonuclease conversion, and E. coli DNA contamination below 10 copies per 2000 U tested. One unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of 32P-ATP in 30 minutes at 37°C in 1X T4 Polynucleotide Kinase Reaction Buffer.

The enzyme is supplied at a concentration of 10,000 U/mL in 10 mM Tris-HCl, 50 mM KCl, 0.1 µM ATP, 1 mM DTT, 0.1 mM EDTA, 50% glycerol (pH 7.4 at 25°C) and is accompanied by a 10X reaction buffer containing 700 mM Tris-HCl, 100 mM MgCl2, and 50 mM DTT (pH 7.6 at 25°C). Recommended storage temperature is -25°C to -15°C, and the product is developed, manufactured, and sold for in vitro use only.