DNA Ligase (E. coli) is a typical DNA ligase that catalyses the formation of a phosphodiester bond between the 5´-phosphate and the 3´-hydroxyl of two adjacent DNA strands in duplex DNA. Thus, the enzyme is particularly effective to repair nicks in dsDNA although it can be used to ligate two DNA fragments with compatible cohesive ends. DNA Ligase (E. coli) is not significantly active on blunt-ended substrates. DNA Ligase (E. coli) uses NAD as a cofactor and can be heat-inactivated. The enzyme is active at a range of temperatures (4 °C – 37 °C).
Applications:
– Gene cloning
– Second-strand cDNA synthesis
– Phosphodiester bond ligations