T4 PNK Highlights

T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer of the terminal gamma-phosphate from ATP to the 5′-OH group of double- and single-stranded DNA, RNA and nucleoside 3′-monophosphate molecules. T4 PNK also exhibits 3′-phosphatase activity and 5′-ADP phosphatase activity.

  • Prepare DNA for ligation through 5’ DNA phosphorylation and 3’ DNA dephosphorylation
  • Reduce material costs with a drop-in solution that delivers equivalent performance at a differentiated price-point
  • Custom formats available, including high concentration
  • Highly stringent enzyme manufacturing ensures quality performance across lots


  • NGS library preparation
  • Cloning (5′-phosphorylation of DNA and RNA prior to ligation)
  • Removal of 3′-phosphates
  • 5′-labeling of DNA and RNA to be used as probes, primers, and markers


High-purity T4 DNA Ligase

Available with standard and rapid ligation buffers for simple optimization


QC Specifications
Description Specification
Protein Purity Assay ≥ 99%
dsDNA Exonuclease Assay* <1% released
ssDNA Exonuclease Assay* <1% released
DNA contamination Assay (E. coli, mammalian, library)* < 10 copies
Phosphatase Contamination Assay* < 1% released
Endonuclease Contamination Assay* Not detectable

*As assessed using 250 U of enzyme input per assay.


Unit definition: One unit of T4 Polynucleotide Kinase is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of phosphate onto a DNA substrate from an ATP donor in 30 minutes at 37°C

Reaction conditions:
1X T4 PNK Reaction Buffer
Incubate at 37°C

Storage buffer: 10mM Tris-HCl, pH7.4, 50mM KCl, 0.1 EDTA, 1mM DTT, 50% Glycerol, 0.1 µM ATP
1X T4 PNK Reaction Buffer: 70 mM Tris-HCl, pH 7.6 at 25°C, 10 mM MgCl₂, 5 mM DTT

Heat inactivation: 65°C for 20 min

Molecular weight: 34.6 kDa