E. coli DNA ligase catalyzes the phosphodiester bond formation between an adjacent 5′ phosphate and a 3′ hydroxyl of DNA ends, requiring NAD+ and Mg2+ as cofactors. Ligation of blunt ended DNA is extremely inefficient relative to cohesive DNA end ligation and nick sealing.
Source of ProteinThe gene encoding E. coli DNA Ligase expressed from a plasmid in E. coli.
Supplied in10 mM Tris-HCl50 mM KCl1 mM DTT0.1 mM EDTA50% glycerolpH 7.5 @ 25°C
Supplied With:B6090 (10X E.coli DNA Ligase Reaction Buffer)
10X E.coli DNA Ligase Reaction Buffer (B6090):300 mM Tris-HCl40 mM MgCl2260 µM NAD10 mM DTT0.5 mg/mL BSApH 8.0 @ 25°C
Unit Definition1 unit is defined as the amount of E.coli DNA Ligase required to ligate 50% of 100 ng DNA fragments with cohesive termini in 30 minutes at 25°C.
