T4 DNA Ligase catalyzes the formation of phosphodiester bonds in the presence of ATP between double-stranded DNAs with 3'hydroxyl and 5'phosphate termini. Single-stranded nucleic acids are not substrates for this enzyme.
Optimizes ligation in 5 minutes
Source: Purified from E. coli lambda lysogen NM989
Performance and quality testing: Endodeoxyribonuclease, 3' and 5' exodeoxyribonuclease assays; ligation efficiency tested
Unit definition: One unit catalyzes exchange of 1nmol 32P-labeled pyrophosphate into ATP in 20 min. at 37°C. One unit is equal to approximately 300 cohesive-end ligation units
Unit reaction conditions: 66mM Tris-HCl (pH 7.6), 6.6mM MgCl2, 10mM DTT, 66μM ATP, 3.3μM 32P-labeled pyrophosphate, and enzyme in 0.1mL for 20 min. at 37°C