E. coli DNA Ligase

Source

Escherichia coli

Reagents Supplied

5X E. coli Reaction Buffer

Applications

• Catalyzes the formation of a phosphodiester bond between duplex DNA fragments with cohesive ends
• Suitable for high-efficiency cloning of full-length cDNA (1,2)
• Condensation of the 5'-phosphoryl group with adjacent 3'-hydroxyl group is coupled with the hydrolysis of NAD+

Unit Definition

One unit is the amount of enzyme required to yield 50% ligation of HindIII fragments of lambda DNA in 30 minutes at 16°C in a 20 µl assay mixture containing a DNA terminus concentration of 0.12 µM (300 µg/ml).

Storage Buffer

10 mM Tris-HCl (pH 7.4)
50 mM KCl
0.1 mM EDTA
10 mM Ammonium Sulfate
1 mM Dithiothreitol 
50% (v/v) Glycerol

Storage Conditions

Store at -20°C
Product shipped on dry ice