|Source||Isolated from E.coli strain that carries the cloned Polynucleotide kinase gene from bacteriophage T4|
|Description||T4 polynucleotide Kinase 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. The enzyme also catalyzes the removal of 3 - phosphoryl groups from 3 - phosphoril polynucleotides, deoxynucleoside 3 - monophosphates and deoxynucleoside 3 - diphosphates.|
- end-labeling DNA or RNA for probes and DNA sequencing;
- addition of 5 - phosphates to oligonucleotides to allow subsequent ligation;
- removal of 3 - phosphoril groups.
|Unit definition||One unit is the amount of enzyme catalyzing the incorporation of 1 nmol of acid-insoluble [32P] in 30 minutes at 37°C.|
Unit Assay Conditions: 1 x T4 polynucleotide kinase Buffer, 66 µM [γ- 32P] ATP (5 x 106 cpm/µmol) and 0.26 mM 5 - hydroxyl - terminated salmon sperm DNA.
|Reaction buffer||SE-buffer polynucleotide kinase T4, (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 5 mM DTT.)|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 50% glycerol; Store at -20°C.|
|Quality control||The enzyme is purified free of contaminating endonucleases, exonucleases, phosphatases. |
|MSDS:||Download MSDS as PDF|