Thermo Scientific™ T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer of gamma-phosphate from ATP to single-stranded and 5'-OH group of double-stranded DNA and RNA, oligonucleotides or nucleoside 3'-monophosphates (forward reaction). This reaction is reversible. In the presence of ADP, T4 polynucleotide kinase exhibits 5'-phosphatase activity and catalyzes the exchange of phosphate groups (exchange reaction) between 5'-P-oligonucleotide-polynucleotide and ATP. This enzyme is also a 3'-phosphatase.

Product Advantages
• Restriction enzymes, RT and T4 DNA LigaseActive in all buffers

Applications
• Label the 5'-end of nucleic acids for use in:< br> --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 before ligation
• Phosphorylation of PCR primers
• By [³² P]-DNA modification detection by post-labeling assay
• Removal of 3'-phosphate group

Note
• Can be through forward reaction or exchange The reaction labels the 5'-end of the nucleic acid.
• Polyethylene glycol (PEG) and spermidine improve the rate and efficiency of phosphorylation reactions. PEG was used to exchange the reaction mixture.
•Since T4 polynucleotide kinase is inhibited by ammonium ions, use sodium acetate to precipitate DNA before phosphorylation.