T4 Polynucleotide Kinase (10 U/µL)

Catalog number: EK0031

Thermo Scientific™  Related applications: Restriction Enzyme Cloning

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Description

Thermo Scientific T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer of the gamma-phosphate from ATP to the 5'-OH group of single- and double-stranded DNAs and RNAs, oligonucleotides or nucleoside 3'-monophosphates (forward reaction). The reaction is reversible. In the presence of ADP T4 Polynucleotide Kinase exhibits 5'-phosphatase activity and catalyzes the exchange of phosphate groups between 5'-P-oligo-polynucleotides and ATP (exchange reaction).

The enzyme is also a 3'-phosphatase.

Highlights

Active in Thermo Scientific restriction enzyme, RT, and T4 DNA Ligase buffers

Applications

• probes for hybridization
• probes for transcript mapping
• markers for gel electrophoresis
• primers for DNA sequencing
• primers for PCR

• 5'-phosphorylation of oligonucleotide, PCR products, other DNA or RNA prior to ligation
• Phosphorylation of PCR primers
• Detection of DNA modification by the [32P]-postlabeling assay• Removal of 3'-phosphate groups

Notes

Polyethylene glycol (PEG) and spermidine improve the rate and efficiency of the phosphorylation reaction. PEG is used in the exchange reaction mixture.

As T4 Polynucleotide Kinase is inhibited by ammonium ions, use sodium acetate to precipitate DNA prior to phosphorylation.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Enzyme: T4 Polynucleotide Kinase
Product Size: 500 units

Documents

Manuals & protocols

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