NEB, M0238S, 9°N™ DNA Ligase

[{"group": "General", "name": "Unit Definition (Cohesive End Unit) One unit is defined as the amount of enzyme required to give 50% ligation of the 12-base pair cohesive ends of 1 µg of BstEII-digested λ DNA in a total reaction volume of 50 μl in 15 minutes at 45°C. A cohesive end unit is equivalent to the nick-closing unit (1). Reaction Conditions 1X 9°N DNA Ligase Reaction Buffer Incubate at 45°C 1X 9°N DNA Ligase Reaction Buffer 10 mM Tris-HCl 600 µM ATP 2.5 mM DTT 2.5 mM MgCl2 0.1% Triton® X-100 (pH 7.5 @ 25°C) Usage Concentration 40,000 units/ml Storage Buffer 10 mM Tris-HCl 50 mM KCl 1 mM DTT 0.1 mM EDTA 200 µg/ml Recombinant Albumin 50% Glycerol 10 mM ammonium sulfate pH 7.4 @ 25°C Heat Inactivation No Unit Assay Conditions 1X 9°N DNA Ligase Reaction Buffer and 20 µg/ml BstEII-digested λ DNA in a 50 μl reaction. After incubation at 45°C for 15 minutes, the reaction is terminated by addition of stop dye (50% glycerol, 50 mM EDTA and bromophenol blue), heated at 70°C for 10 minutes and then loaded on a 0.7% agarose gel. Due to the presence of ligase, the cos ends of BstEII-digested λ DNA will stay together after 70°C heat treatment.", "value": ""}]