T4 Polynucleotide Kinase catalyzes the transfer of the g-phosphate from ATP to the 5´-terminus of polynucleotides or to mononucleotides bearing a 5´-hydroxyl group.
The enzyme can be used to phosphorylate RNA, DNA and synthetic oligonucleotides.T4 Polynucleotide Kinase is purified from a recombinant E. coli strain.Concentration & Storage Condition- 10 unit/㎕. Store at -20℃.Storage Buffer- 20 mM Tris-HCl, pH 7.5, 25 mM KCl, 2 mM DTT, 0.1 mM EDTA, 0.1 mM ATP and 50% glycerol.10x Reaction Buffer- 700 mM Tris-HCl, pH 7.6, 100 mM MgCl2, 50 mM DTT.Unit Definition- One unit is defined as the amount of enzyme required to catalyze the transfer of 1 nmole of phosphate to the 5´-OH end of a polynucleotide from [g-32P]ATP in 30 min at 37°C. The reaction conditions are: 40 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 5 mM DTT, 0.1 mM [g-32P] ATP and 0.5 mM 5´-OH polynucleotide end concentration.QC Tests
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.