T4 DNA Ligase 50-200u/µl

Description T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 #39;-phosphate and 3 #39;-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. Concentrations 50 - 200u/μl Features Ultrapure recombinant protein Seals single-stranded nicks in duplex DNA, RNA or DNA-RNA hybrids. ATP is an essential cofactor for the reaction. Supplied With 10X Buffer T4 Ligase 50mM Tris-HCl (pH7.8 at 25˚C), 10mM MgCl2, 10mM DTT, 1mM ATP and 25μg/ml BSA. Store at -20˚C. Storage Buffer 10mM Tris-HCl (pH7.5), 50mM NaCl, 0.1mM EDTA, 10mM 2-mercaptoethanol and 50% glycerol. Store at -20˚C. Thermal Inactivation 65˚C for 15 minutes Unit Definition 1u (*Cohesive End Ligation Unit) is defined as the amount of enzyme that is required to give 50% ligation of Hind III fragments of lambda DNA (5 #39; DNA termini concentration of 0.12μM [300μg/ml]) in 20μl of 1X T4 DNA Ligase Buffer in 30 minutes at 16˚C. *One Cohesive End Ligation Unit is equal to 0.015 Weiss units. Equivalently, one Weiss unit is equal to 67 Cohesive End Ligation Units. Application Catalyzes the linkage of 5 #39; or 3 #39; blunt/cohesive ends of double-stranded DNA by formation of phosphodiester bond. Joining of oligonucleotide linkers or adapters to blunt ends. Repair nicks formation in duplex nucleic acids. Quality Control All preparations are assayed for contaminating endonuclease, exonuclease and non-specific DNase activities.