T4 DNA Recombinant eiwit

Source : Escherichia Colilambda lysogen NM 989. 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.

DNA ligase 4||EC 6.5.1.1||DNA ligase IV||Polydeoxyribonucleotide synthase [ATP] 4.

50mM KCl, 10mM Tris-HCl (pH 7.4), 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA and 50% glycerol.

50mM KCl, 10mM Tris-HCl (pH 7.4), 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA and 50% glycerol. Store at -20C. 50mM KCl, 10mM Tris-HCl (pH 7.4), 0.1mM EDTA, 1mM DTT, 200 μg/ml BSA and 50% glycerol. Store at -20C.

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 > 200mM.• 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 50μM.• To dilute T4 DNA Ligase that will subsequently be stored at –20°C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used.