E. coli DNA Ligase

Description:
E. coli DNA Ligase is a NAD⁺-dependent enzyme that catalyzes the formation of phosphodiester bonds between complementary 3'-hydroxyl and 5'-phosphoryl termini of dsDNA. The enzyme works best with cohesive dsDNA ends and is also active on nicked DNA. Blunt ends can be ligated in the presence of condensing reagents such as polyethylene glycol or Ficoll^®. A 10x Reaction Buffer is provided with the enzyme.

Source:
E. coli strain containing an overproducing clone of E. coli DNA Ligase.

Application:
- Ligation for cloning
- Okayama and Berg cDNA cloning

Unit Definition:
One unit of E. coli DNA Ligase is defined as the amount of enzyme required to provide ligation (>50%) of Hind III digested phage lambda DNA (5'-DNA termini concentration of 0.12 µM, 300 µg/ml) in 30 minutes at 16°C under standard assay conditions.

Reaction Conditions:
10x E. coli DNA Ligase Reaction Buffer
Incubate at 16°C

10x E. coli DNA Ligase Reaction Buffer:
300 mM Tris-HCl
40 mM MgCl₂
260 µM NAD
10 mM DTT
pH 8.0 @ 25°C

Specific Activity: 10,000 Unit/mg

Recommended Storage Condition: -20°C