| 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.
Applications:
- 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.
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| Unit | 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.
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| Reaction buffer | SE-buffer polynucleotide kinase T4, (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 5 mM DTT.) |
| Optimal temperature | 37oC |
| 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 | Free of exonuclease, phosphatase, endonuclease and RNase activities. |
| MSDS: | Download MSDS as PDF |
