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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
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Applications
- 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
ALSO AVAILABLE
High-purity T4 DNA Ligase
Available with standard and rapid ligation buffers for simple optimization
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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.
Properties
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