T4 DNA Ligase catalyzes the joining of two strands of DNA between the 5'-phosphate and the 3'-hydroxyl groups. DNA fragments may have either blunt or cohesive ends. T4 DNA Ligase catalyzes the joining of RNA to either DNA or RNA strands in a duplex molecule, but will not join single-stranded nucleic acids. Unit Definition: One Weiss unit is defined as the amount of enzyme required to catalyse the conversion of 1 nmole of 32P-labelled pyrophosphate into Norit-adsorbable material in 20 minutes at 37 C. 0.014 Weiss units of ligase is the equivalent of 1 ligation unit which catalyzes greater than 95% ligation of 1ug of Lambda/Hind III fragments at 16C in 20 minutes. The reaction conditions for Weiss unit definition are as follows: 66mM Tris-HCl, pH 7.6, 6mM MgCl2, 1mM DTT, 0.066mM ATP and 3.3uM 32PPi . Note: 1 Weiss Unit = approx 67 cohesive end units.
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