T4 DNA ligase can catalyze the formation of a phosphodiester bond between juxtaposed 5' phosphate and 3' hydroxyl termini in duplex DNA or RNA. ATP is required as a coenzyme to catalyze not only the connection of the blunt end and cohesive end termini but also the repair of single-stranded nicks in duplex DNA and some DNA/RNA hybrids.
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Cat. No.: T4DL-40k (for 40KU)
Cat. No.: T4DL-200k (for 200KU)
One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (5´ DNA termini concentration of 0.12 µM, 300- µg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C.
1. Set up the following reaction in a microcentrifuge tube on ice
2. For cohesive (sticky) ends, incubate at 16°C for 3 hours or 4°C overnight.
The minimum shelf life is 1 year at -20°C and 3 years at -80°C.