T4 DNA Ligase

T4 DNA Ligase catalyzes the joining of two cohesive- or blunt-ended strands of DNA between the 5´-phosphate and the 3´-hydroxyl groups of adjacent nucleotides. The enzyme will not join single-stranded nucleic acids. T4 DNA Ligase is provided with 10X Reaction Buffer: 300mM Tris-HCl (pH 7.8 at 25°C), 100mM MgCl₂, 100mM DTT and 10mM ATP.

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Reference

1. Engler, M.J. and Richardson, C.C. (1982) In: The Enzymes, Boyer, P.D., ed., Academic Press, New York, NY.

Note: Units listed are Weiss units.

Storage Buffer: 10mM Tris-HCl (pH 7.4 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA and 50% glycerol.

Source: Recombinant E. coli strain.

QC Tests: Activity, SDS-PAGE/purity, DNase, endonuclease, RNase, blue/white cloning assay.

Unit Definition: 0.01 Weiss unit of T4 DNA Ligase is the amount of enzyme required to catalyze the ligation of greater than 95% of 1μg of λ/HindIII fragments at 16°C in 20 minutes.