E. coli DNA ligase can catalyze the formation of phosphodiester bonds between the 5’-PO4 ends and 3’-OH ends of adjacent DNA strands. It exhibits almost no detectable activity for blunt-end substrate DNA. The optimal pH for this reaction is 7.5-8.0 (Tris-HCl buffer), and NAD is required as a coenzyme. Unlike T4 DNA Ligase, which can join both sticky and blunt ends, this enzyme only catalyzes the ligation between cohesive-ended DNA molecules. However, in the presence of PEG and high concentrations of monovalent cations, it can also ligate blunt-ended DNA. Additionally, this product cannot catalyze the ligations between DNA and RNA or between RNA molecules.This enzyme can be used for ligation reactions in NGS methylation library and can also be employed in the Okayama-Berg method for cDNA cloning. Specifications Concentration 60 U/μL Unit Definition In a 20 μL ligation reaction system, the amount of enzyme required to ligate over 90% of 6 μg of λ DNA-Hind III fragments at 16°C for 30 minutes is defined as one unit of activity (U) Components Components No. Name 14955ES82 14955-A E. coli DNA ligase(60 U/μL) 20 μL 14955-B 10× E. coli DNA ligase Buffer 100 μL 14955-C 10× BSA(0.05%) 100 μL Note: BSA should be protected from repeated freezing and thawing. For short-term storage, it can be kept at 4℃.
