Recombinant T4 DNA Ligase

Biomolecules

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5' -phosphate and 3' -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

One Weiss unit is equivalent to circa 67 cohesive-end ligation units. T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200 mM. • Ligation of blunt-ended and single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 (10% w/v final concentration) or hexamine chloride, or by reducing the ATP concentration to 50 uM. • To dilute T4 DNA Ligase that will subsequently be stored at -20C, 50% glycerol storage buffer should be used, to dilute for immediate use, 1 x T4 DNA Ligase reaction buffer can be used.