Thermo Fisher, 18052019, E. coli DNA Ligase

Thermo Fisher, 18052019, E. coli DNA Ligase E. coliDNA Ligase catalyzes the formation of phosphodiester bonds in the presence of β-NAD between double-stranded DNAs with 3´ hydroxyl and 5´ phosphate cohesive termini. Single-stranded nucleic acids are not substrates for this enzyme.
Applications:Second-strand cDNA synthesis (1). T4 DNA Ligase alternative when blunt-end ligation is not required (2).
Source:Purified fromE. coli594(Su-) bearing λ lysogen gt4lop-11lig+S7 (3).
Performance and Quality Testing:Endodeoxyribonuclease and 3´ and 5´ exodeoxyribonuclease assays; ligation efficiency tested.
Unit Definition:One unit is the amount of enzyme required to give 50% ligation ofHind III-digested λ DNA in 30 min. at 16°C in a final volume of 20 μl containing a 5´ termini concentration of 0.
12 ´M (300 μg/ml).
Unit Reaction Conditions:18.
8 mM Tris-HCl (pH 8.
3), 90.
6 mM KCl, 4.
6 mM MgCl2, 3.
8 mM DTT, 0.
15 mM λ-NAD, 10 mM (NH4)2SO4in 20 μl for 1 h at 16°C.
Compatible Buffer: 10X Reaction Buffer
Product Type: DNA Ligase
Quantity: 100 U
Shipping Condition: Approved for shipment on Wet or Dry Ice
Enzyme: Ligase
Unit Size: Each