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T4PNK-RO Roche

T4 Polynucleotide Kinase, 3′-phosphatase free

from phage T4 am N81 pse T1 infected Escherichia coli BB

Synonym: kinase, t4 polynucleotide, 3′-phosphatase-free

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Properties

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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)
mfr. no.   Roche
shipped in   dry ice
storage temp.   −20°C

Description

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.

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.

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

Other Notes

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

Kit component only

Description

Product #

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Enzyme Solution, in buffer, pH 7.5 (+25 °C) 10 U/μl    
Phosphorylation Buffer 10x concentrated    
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

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Protocols & Articles
Peer-Reviewed Papers
15

References

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