Suitable for high-efficiency cloning of full-length cDNA (1, 2).
Unit Definition: One unit is defined as the amount of enzyme required to yield 50% ligation of Hind III fragments of lambda DNA. Incubation is at 16°C in 20 µl of assay mixture with a DNA terminus concentration of 0.02 µM (50 µg/ml).
Storage Conditions: Store at –20°C.
1 x Reaction Buffer: 30 mM Tris-HCl (pH 8.0 at 22°C), 1 mM dithiothreitol, 4 mM MgCl2, 26 µM NAD+, 50 µg/ml bovine serum albumin.
Optimal ligation occurs at 16°C.
Storage Buffer: 10 mM Tris-HCl (pH 7.4 at 22°C), 50 mM KCl, 0.1 mM EDTA, 10 mM ammonium sulfate, 1 mM dithiothreitol and 50% (v/v) glycerol.
Quality Control: All preparations are tested for contaminating endonuclease and exonuclease activities, along with functional testing in the ligation reaction.
Ligation Assay Conditions: 30 mM Tris-HCl (pH 8.0 at 22°C), 4 mM MgCl2, 1.2 mM EDTA, 1 mM dithiothreitol, 0.026 mM NAD+, 50 µg/ml bovine serum albumin and Hind III fragments of lambda DNA. Incubation is at 16°C for 30 min.
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