Description

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5’-phosphate and 3’-hydroxyl termini in duplex DNA or RNA. This enzymes joins DNA fragments with either cohesive or blunt termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

Product Includes

  • T4 DNA Ligase
  • 10x T4 DNA Ligase Reaction Buffer

Purity: ≥99% (assessed by SDS-PAGE with Coomassie blue staining)

Concentration: 10 unit/μlOne unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (250 ng/µl) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X T4 DNA ligase reaction buffer.

Source: E. coli strain expressing a recombinant clone

Inhibition and Inactivation:Inhibitors: metal chelators, phosphate and ammonium ions, KCl and NaCl at a concentration higher than 50 mM.Inactivated by heating at 70°C for 15 min or by addition of EDTA.

Quality ControlEndonuclease Activity (Nicking):1 µg of supercoiled plasmid DNA is incubated with 20 units of T4 DNA Ligase in 1x Ligase buffer for 2 hours at 37 ºC. Following incubation, the supercoiled DNA is visualized on an ethidium bromide stained agarose gel. No visible nicking or cutting of DNA was found.

Storage/Handling: Store at -20°C. Product may be shipped on blue ice.