Storage Condition

-20°C

Components

• T4 Polynucleotide Kinase (10U/μl)
• 10x T4 PNK Reaction Buffer

Description

T4 polynucleotide kinase is short for T4 PNK. It is expressed by E. coli. The source of the expressed gene is T4 bacteriophage, which can catalyze phosphate in ATP γ- It can transfer and exchange with the 5 ‘- hydroxy end of the oligonucleotide chain (double stranded or single stranded DNA or RNA) and the 3’ – monophosphate nucleoside. At the same time, it also has 3 ‘phosphatase activity, which can hydrolyze the 3’ – phosphate group from the 3 ‘phosphate end of the oligonucleotide, deoxy 3’ – monophosphate nucleoside and deoxy 3 ‘- diphosphate nucleoside. The T4 polynucleotide kinase can be used for 5 ‘terminal labeling or phosphorylation of oligonucleotides, DNA or RNA; 5 ‘phosphorylation of mononucleotides that catalyze 3’ phosphorylation and removal of 3 ‘terminal phosphate groups. Heating this product at 75°C for 10 minutes can inactivate it, and adding EDTA can also inactivate it. Metal ion chelating agent, phosphate, ammonium ion, KCl greater than 50 mm and NaCl can significantly inhibit its activity.

Unit Definition

At 37°C, within 30 minutes, the amount of enzyme required for the transfer of 1 nmol of γ- Phosphate group on ATP to the end of 5′-OH on DNA is defined as 1 active unit.

Quality Control

After multiple column purification, the purity of SDS-PAGE was more than 99%; No contamination of endonuclease, exonuclease, phosphatase and RNase activities was detected.

Precautions

• Because the ammonium salt can strongly inhibit the activity of T4 polynucleotide kinase, the DNA obtained from ammonium salt precipitation cannot be used for the labeling reaction of T4 polynucleotide kinase.
• PEG can promote the rate and efficiency of phosphorylation reaction, and PEG should be added to the exchange reaction system.
• When using, the enzyme should be stored in an ice box or on an ice bath, and should be stored at -20°C immediately after use.

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