- 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. The ligation will occur only if the oligonucleotides are perfectly paired to the complementary target DNA and have no gaps between them; therefore, a single-base substitution can be detected. This product is active at elevated temperatures (45°C-70°C) (1, 2). The physical purity of this enzyme is ≥99% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below). Product Source E. coli strain expressing the cloned Taq DNA ligase gene from Thermus aquaticus HB8 Applications Allele-specific gene detection by using Ligase Detection Reaction (LDR) and Ligase Chain Reaction (LCR) (1). Mutagenesis by incorporation of a phosphorylated oligonucleotide during primer extension amplification(3). Product Includes 1) Taq DNA Ligase 2) 10X Taq DNA Ligase Buffer with NAD Storage Temperature –20 °C Storage Buffer 50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 ºC 10X Taq DNA reaction buffer with NAD 500 mM Tris-HCl, 100 mM MgCl2, 100 mM DTT, 10 mM NAD , pH 7.5 @ 25ºC Unit Definition One unit is defined as the amount of Taq DNA required to join 50% of 1 μg of the 12-base cohesive ends of Lambda DNA cut with Sma I and Sal I in 50 μl reaction in 15 min incubation at 45 °C. Quality Control This product is free from detectable RNase or contaminating DNA endonuclease activities.3218 3219
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