- Description Intact Genomics (IG®) Tth DNA Ligase is an engineered high-fidelity DNA ligase. It catalyzes the formation of a phosphodiester bond in duplex DNA containing adjacent 5′-phosphoryl and 3′-hydroxyl termini, using NAD as a cofactor. Ligation occurs only when the oligonucleotides are perfectly paired to the complementary target DNA and have no gaps between them; allowing detection of single-base substitutions. IG® high-fidelity Tth DNA Ligase is active at elevated temperatures (45°C–70°C), which gives it clear advantages over standard ligases such as T4 DNA Ligase. It is known for its high fidelity and strong mismatch discrimination, offering better specificity than mesophilic ligases, including Taq DNA Ligase2-3 Product Source: Tth DNA ligase is a derivatized enzyme that originated from Thermus thermophilus. Species: About seventy-seven (~77) kDa DNA ligase originating from Thermus thermophilus with enhanced fidelity. Benefits: Fast: Completes standard ligations in 5 minutes and most applications in under 30 minutes Efficient and High-Fidelity: Delivers higher ligation fidelity than both Taq DNA ligase and T4 DNA ligase, resulting in more accurate and expected products Thermostable: Maintains strong activity at elevated temperatures(45°C–70°C) Highly Specific: Excellent for SNP discrimination and other precision applications Reduces Background Noise: Improves assay accuracy and reliability Buffer Tolerant: Less inhibition by analytes, enabling simplified workflows without buffer dilution, buffer exchange, or purification Applications: Ligase Chain Reaction (LCR) DNA mutation detection Molecular diagnostics and pathogen detection Padlock probe assays and rolling circle amplification (RCA) High-temperature DNA ligation of complex or GC-rich templates Synthetic biology applications Product Components: Enzyme: Tth DNA Ligase Buffer: 10x Ligase Reaction Buffer with NAD Storage and Shipping Conditions: Tth DNA Ligase: Store at -20 °C 10x Ligase Reaction Buffer with NAD : Store at -20 °C Shipping condition: Ship on dry ice or with ice packs Quality Control: Ligase Activity: Tth DNA Ligase can seal nicks in DNA plasmid at ≥74% efficiency and works equal to or better than competitor’s Tth Ligase. For sealing efficiency, IG’s T5 Exonuclease (product catalog #3236/8) digests virtually 100% of nicked DNA but leaves sealed DNA intact to quantify the amount of ligated product. Nuclease Free: Nuclease activity is measured in a 10 μL reaction containing 0.5 μg of plasmid DNA and 0.5 μL of enzyme solution incubated for at least 14 hours at 37 °C. Protein Purity: Proteins are evaluated by SDS-PAGE and quantified to be ≥95% pure.
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