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.
|Product Source||An E. coli strain that carries the cloned T4 Polynucleotide Kinase gene.|
|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|
|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.|