T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5´-phosphate and 3´-hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end and cohesive end termini as well as repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. T4 DNA Ligase is purified from a recombinant E.coli strain.
Reaction Conditions- Incubate at 16-37°C in 1x T4 DNA ligase reaction buffer. 5x T4 DNA Ligase Reaction Buffer- 150 mM Tris-HCl, pH 7.8, 50 mM MgCl2, 5 mM ATP, 50 mM DTT. Unit Definition- 0.01 Weiss unit of enzyme is defined as the amount of enzyme required to give 90% ligation of Hind lll fragments of lambda DNA in 30 min at 16°C in 20 ㎕ of the assay mixture. - 2 unit/㎕. Store at -20℃.
Concentration & Storage Condition
Storage Buffer- 10 mM Tris-HCl, pH 7.4, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol QC Tests
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests