T4 Polynucleotide Kinase, 3′-phosphatase free
ROCHE/T4PNK-RO - from phage T4 am N81 pse T1 infected Escherichia coli BB
Synonym: kinase, t4 polynucleotide, 3′-phosphatase-free
Product Type: Chemical
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 1,000 U (10838292001) |
| pkg of 200 U (10709557001) | |
| Quality Level | 100  |
| specific activity | ≥40 x 103 units/mg protein |
| 40000 U/mg | |
| storage temp. | −20°C |
| Application: | Use T4 Polynucleotide Kinase, 3′-phosphatase-free, to: • Phosphorylate the 5′ ends of RNA or DNA • Phosphorylate the 3′ ends of RNA to prevent cyclization • Add a 5′[32P]-terminal label to 3′-CMP, forming 5′[32P]pCp. This substrate is commonly used for 3′-end labeling of RNA with T4 RNA ligase. • Label 5′-hydroxyl ends of DNA with [?-32P]-ATP (either by direct phosphorylation or by phosphate exchange) • Phosphorylate the 5′-hydroxyl ends of oligonucleotides obtained from automated synthesizers • Phosphorylate the 5′-hydroxyl ends of linkers. • Purify recombinant RNA in E. coli. Note: The 3′-phosphatase activity of the wild-type kinase is not observed under optimal incubation conditions with the mutant T4 Polynucleotide Kinase. |
| Biochem/physiol Actions: | T4 polynucleotide kinase catalyzes the transfer of the terminal phosphate group of ATP to the 5′-hydroxyl terminus of DNA or RNA. It also can catalyze the exchange of 5′-terminal phosphate groups. In contrast to the wild type E. coli enzyme, this phage derivative lacks 3′-phosphatase activity. The assay time is 30 minutes and volume activity is 10000 U/ml. |
| General description: | T4 polynucleotide kinase catalyzes the transfer of the terminal phosphate group of ATP to the 5′-hydroxyl terminus of DNA or RNA. It also can catalyze the exchange of 5′-terminal phosphate groups. In contrast to the wild type E. coli enzyme, this phage derivative lacks 3′-phosphatase activity. The assay time is 30 minutes and volume activity is 10000 U/ml. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Packaging: | 1 kit containing 2 components |
| Quality: | Absence of 3′-phosphatase, endonucleases, nicking activity, 3′-exonucleases, and ribonucleases, tested according to the current Quality Control procedures; function tested using the DNA 5′-End Labeling Kit. |
| Unit Definition: | One unit is the enzyme activity which catalyzes the incorporation of 1 nmol 32P into acid-precipitable products within 30 minutes at +37 °C. Volume Activity: 10 x 103 U/ml |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| activity | specific activity: ≥40 x 103 units/mg protein; specific activity: 40000 U/mg |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |
| Components | Enzyme Solution, in buffer, pH 7.5 (+25 °C) 10 U/μl; Phosphorylation Buffer 10x concentrated |