One Weiss unit of T4 DNA Ligase converts 1 nmole of p32 from pyrophosphate into Norit-adsorbable material in 20 minutes at 37℃ in 33 mM Tris-acetate (pH 7.8), 66 mM potassium acetate, 10 mM magnesium acetate, 0.5 mM DTT, and 1 mM ATP. One Weiss unit equals approximately 200 cohesive- end ligation units.
T4 DNA Ligase is functionally tested in cloning assays and is free of detectable contaminating DNA exo- and endo-nuclease and RNase activities.
1 Weiss unit/㎕
50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 0.1% Triton X-100, and 1 mM DTT, 50% Glycerol
10X Reaction buffer
composed of 400mM Tris-HCl, 100mM MgCl2, 100mM DTT, 5mM ATP (pH 7.8 at 25°C).
Cloning of restriction fragments
Joining linkers and adapters to blunt-ended DNA
T4 DNA Ligase is the most versatile and commonly used ligase for DNA cloning. This ATP-dependent enzyme covalently joins 5'-phosphates to 3'-hydroxy-lated termini at the blunt or compatible cohesive ends of double-stranded DNA fragments produced by restriction enzyme digestion or other enzymatic processes.
T4 DNA Ligase has no activity on single- stranded nucleic acids. Following a ligation reaction, T4 DNA Ligase may be inactivated by incubation at 65℃ for 10 minutes.