T4 Polynucleotide Kinase is a 5'-hydroxyl kinase for polyribonucleotides, capable of catalyzing the transfer of the γ-phosphate group from ATP to the 5'-hydroxyl terminus of oligonucleotide chains (double-stranded or single-stranded DNA or RNA) and to 3'-monophosphate nucleotides. This reaction is reversible. T4 Polynucleotide Kinase also possesses 3'-phosphatase activity, hydrolyzing 3'-phosphate groups from the 3'-phosphate terminus of oligonucleotides, deoxy-3'-monophosphate nucleotides, and deoxy-3'-diphosphate nucleotides. In the presence of ADP, T4 PNK exhibits 5'-phosphatase activity, catalyzing the exchange of 5'-phosphate groups between 5'-P-oligo/polyribonucleotides and ATP. It is suitable for DNA library construction and probe labeling. Compared to T4 Polynucleotide Kinase (Cat#12902ES), UCF.ME™T4 Polynucleotide Kinase has lower host residue, making it suitable for applications that require stricter background control, such as pathogen detection.

Applications

Phosphorylation of the 5' terminus of DNA or RNA for ligation reactions.

End labeling of DNA or RNA for probe preparation and DNA sequencing.

Phosphorylation of the 5' terminus of mononucleotides that already have a 3'-phosphate group to prepare pNp substrates for addition to the 3' end of DNA or RNA.

Labeling of the 5' terminus of oligonucleotides that have a 3'-phosphate group.

Components

Figures

Figure 1. Monomer Phosphorylation Test

Note: A 300 bp monomer ligation test was performed using different batches of UCF.ME™ T4 Polynucleotide Kinase (10 U/μL) at 5 and 10 U, compared with Brand A. The gel image results showed that Yeasen's product outperformed Brand A in phosphorylation.