Thermo Fisher, 18052019, <i>E. coli</i> DNA Ligase

Quantity: 100 U
Unit Size: Each
Single-stranded nucleic acids are not substrates for this enzyme.
• Source: Purified fromE. coli594 (Su^-) bearing λ lysogen gt4lop-11lig^+S7
• Performance and Quality Testing: Endodeoxyribonuclease and 3' and 5' exodeoxyribonuclease assays; ligation efficiency tested
• Unit Definition: One unit is amount of enzyme required to give 50% ligation ofHind III-digested λ DNA in 30 min. at 16°C in a final volume of 20μL containing a 5' termini concentration of 0.12 'M (300μg/mL)
• Unit Reaction Conditions: 18.8mM Tris-HCl (pH 8.3), 90.6mM KCl, 4.6mM MgCl_2, 3.8mM DTT, 0.15mM λ-NAD, 10mM (NH_4)_2SO_4in 20μL for 1 hr. at 16°CSecond-strand cDNA synthesis; T4 DNA Ligase alternative when blunt-end ligation is not required; Cloning, Restriction Enzyme Cloning