T4 Polynucleotide Kinase

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Cat. NO. AB-RK20524 (datasheet)

T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer and exchange of Pi from the γ position of ATP to the 5´ -hydroxyl terminus of polynucleotides (double-and single-stranded DNA and RNA) and nucleoside 3´-monophosphates. T4 Polynucleotide Kinase also catalyzes the removal of 3´-phosphoryl groups from 3´-phosphoryl polynucleotides, deoxynucleoside 3´-monophosphates and deoxynucleoside 3´-diphosphates.
T4 PNK is applicable to end-labeling DNA or RNA for probes and DNA sequencing, addition of 5´-phosphates to oligonucleotides to allow subsequent ligation and removal of 3´-phosphoryl groups.

Featured Specification
Product Source An E. coli strain that carries the cloned T4 Polynucleotide Kinase gene.
Applications Phosphorylation (Kinase)
Unit Definition One Richardson unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of acid-insoluble [32P] in a total reaction volume of 50 μl in 30 minutes at 37°C in 1X T4 PNK Reaction Buffer with 66 µM [γ-32P] ATP (5 x 106 cpm/µmol) and 0.26 mM 5´-hydroxyl-terminated salmon sperm DNA.
Storage Conditions 10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, 0.1 uM ATP pH7.4 @ 25°C
Storage Temperature -20°C
Reaction Conditions 1X T4 PNK Reaction Buffer, Incubate at 37°C
Reaction Buffer or Mix 1X T4 PNK Reaction Buffer:
70 mM Tris-HCl, 10 mM MgCl2, 5 mM DTT, pH7.6 @ 25°C
Heat Inactivation 65°C for 20 min.