T4 DNA Ligase - 2000 units/µL
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.
- T4 DNA Ligase
- 10x T4 DNA Ligase Reaction Buffer
Purity: ≥99% (assessed by SDS-PAGE with Coomassie blue staining)
Concentration: 2000 unit/μl
One 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.
Endonuclease 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 37ºC. 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.