T4 Polynucleotide Kinase, 3-phosphatase free

Кат. номер

10838292001

10709557001

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.
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.
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
For life science research only. Not for use in diagnostic procedures.

Quality Level	100,
form	solution
specific activity	40 x 103 units/mg protein, 40000 U/mg
packaging	pkg of 1,000 U (10838292001), pkg of 200 U (10709557001)
Торговая марка	Roche
shipped in	dry ice
storage temp.	20°C

RIDADR

NONH for all modes of transport