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
• Labeling 5' -termini of nucleic acids to be used as:
    --Probes for hybridization
    --Probes for transcript mapping
    --Markers for gel electrophoresis
    --Primers for DNA sequencing
    --Primers for PCR
• 5'-phosphorylation of oligonucleotides, 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
• The 5'-termini of nucleic acids can be labeled by either the forward or the exchange reaction.
• Polyethylene glycol (PEG) and spermidine improve the rate and efficiency of the phosphorylation reaction. PEG is used in the exchange reaction mixture.
• Since 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.

Figures

Specifications

Enzyme: T4 Polynucleotide Kinase
Product Size: 500 units

Documents

Manuals & protocols

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