T4 DNA Ligase (Recombinant)

Source E. Coli. cloned gene-30, bacteriophage T4.

Description Recombinant 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.

Physical Appearance Sterile filtered liquid formulation having a concentration of 400 U/µL.

Formulation T4 DNA Ligase is supplied in 50 mM KCl, 20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, 50% glycerol.

Reaction Conditions 50 mM Tris-HCl (pH 7.5 @ 25°C), 10 mM MgCl2, 10 mM DTT, 1 mM ATP. Incubate at 16°C.

Stability Store T4 DNA Ligase at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Avoid multiple freeze-thaw cycles.

Unit Definition 1U is the amount of T4 DNA Ligase required to ligate > than 50% DNA fragments in a 20μl ligation reaction system, 6μg of λDNA-Hind III decomposition product reacts at 16°C for 30 minutes.

1 Weiss unit = 200 cohesive end units.

Inactivation T4 DNA Ligase can be inhibited by NaCl/KCl at a concentrations >than 200mM or by heating at 65 °C for 10 min or at 70 °C for 5 min.

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