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Product description: Taq DNA ligase catalyzes the formation of a phosphodiester bond in duplex DNA containing adjacent 5'-phosphoryl and 3'-hydroxyl termini, using NAD+ as a cofactor.
Source: Recombinant E.coli
Concentration and Size: 40U/μL
Unit Definition: One unit is defined as the amount of enzyme required to give 50% ligation of the 12-base pair cohesive ends of 1μg of BstEII-digested λDNA in a total reaction volume of 50μL in 15minutes at 45℃.
Storage Buffer: 10mM Tris-HCl (pH 7.5, 25℃), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.1% Triton X-100 and 50% (v/v)Glyercol.
Companion Product: 10X Taq DNA Ligase Reaction Buffer, Cat#ON-72, 200mM Tris-HCl pH 7.6, 25℃, 250mM KAC, 100mM Mg(AC)2, 10mM NAD+,0.1% Triton X-100.
Note: The ligation reaction requires NAD+ as a cofactor. NAD+ is supplied in the 10×Taq DNA Ligase Reaction Buffer, the buffer should be stored at -70℃ to extend the half life of the NAD+ cofactor. Taq DNA Ligase is active at 45-65℃
1. Add the following reagents in a PCR tube:
|Nuclease-free H2O||to 50μL|
|DNA||up to 1μg of DNA|
|10×Taq DNA ligase reaction buffer||5μL|
|Taq DNA ligase||2μL (80 units)|
2. Incubate at 45℃ for 15 min.
Storage Conditions: -20℃
Quality Assurance: Free of endonuclease, exonuclease, and Nickase activities.
Physical Purity: ＞95% by SDS-PAGE.
1. Barany F. Proc Nat Acad Sci USA. 88, 189–193 (1991).
2. Gail Lauer, Edwin A. Rudd, Dianel L. Mckay, et al. Journal Of Bacteriology. 173, 5047-5053(1991).