1st July 2026

T4 Polynucleotide Kinase- Cloned

Applications
  • LabeLling of 5´ termini of DNA and RNA with32P or33P for DNA sequencing, blot-hybridization experiments, or transcript mapping using Mung Bean Nuclease, S1 nuclease, or other nucleases.1,2
  • Phosphorylation of oligonucleotide linkers and other DNA or RNA molecules prior to ligation, or for use in ligation amplification reactions with Ampligase® Thermostable DNA Ligase.
  • Preparation of labeled DNA or RNA molecular weight markers for gel electrophoresis and chromatography.

T4 Polynucleotide Kinase (PNK) catalyzes the transfer of the γ-phosphate from ATP to the 5´ hydroxyl of ssDNA and dsDNA, RNA, and nucleoside 3´ monophosphates. The enzyme also removes the 3´ phosphate from 3´-phosphoryl polynucleotides, deoxyribonucleoside 3´ monophosphates, and deoxyribonucleoside-3´,5´-diphosphates to form a 3´-hydroxyl group.

Unit Definition:
One unit of T4 Polynucleotide Kinase converts 1 nmol of32P from [γ-32P]-ATP into an acid-insoluble form in 30 minutes at 37°C under standard assay conditions.

Storage Buffer:
50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 0.1% Triton® X-100, and 1 mM DTT.

T4 PNK 10X Reaction Buffer:
330 mM Tris-acetate (pH 7.5), 660 mM potassium acetate, 100 mM magnesium acetate, and 5 mM DTT. A 10 mM solution of ATP for nonisotopic applications is available separately.

Quality Control:
T4 PNK is tested in 5´ phosphorylation of nucleic acids and is free of detectable exo- and endonuclease and RNase activities.

References

  1. Sambrook, J.et al.(1989) in:Molecular Cloning: A Laboratory Manual(2nd ed.), Cold Spring Harbor Laboratory Press, New York.
  2. Chaconas, G.et al.(1980)Meth. Enzymol.65, 75.

Figure 1. The absence of DNA exonuclease activity in Epicentre’s T4 Polynucleotide Kinase.40-mer oligonucleotides with (5´-PO4) or without (5´-OH) phosphorylated 5´-termini were incubated with the indicated number of units of T4 PNK in 1X T4 PNK Reaction Buffer for 16 hours at 37°C. Electrophoresis of the incubation mixtures in a 15% polyacrylamide/8 M urea gel demonstrates no degradation of the oligonucleotides.

 

Ordering

Catalog No. Concentration Size
T4 Polynucleotide Kinase, Cloned
P0503K                     10 U/µl 3,000 Units

Each T4 Polynucleotide Kinase Kit includes T4 Polynucleotide Kinase at 10 U/µL and 10X Reaction Buffer without ATP. ATP is available separately.

The “enzyme only” format contains T4 Polynucleotide Kinase at 10 U/µL.