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.
Purified from a recombinant E. coli strain.
• Cloning of restriction fragments
• Joining linkers and adapters to blunt-ended DNA
One unit is the amount of enzyme required to yield 50% ligation of HindIII fragments of lambda DNA in 30 min at 16 °C in a 20 μl assay mixture containing a DNA terminus concentration of 0.12 μM.
1×T4 DNA Ligase Reaction Buffer [50 mM Tris-HCl (pH 7.5 at 25°C), 10 mM MgCl2, 10 mM DTT, 1 mM ATP; incubate at 16°C.