Highly specific thermostable DNA ligase for high-temperature applications

• Assemble complex DNA constructs using Gibson cloning methods
• Detect SNPs with high specificity and sensitivity
• Identify expanded trinucleotide repeats using repeat expansion detection (RED)

Ampligase® Thermostable DNA Ligase is a DNA ligase derived from a thermophilic bacterium

Ampligase is stable and active at much higher temperatures than conventional DNA ligases. The half-life of the enzyme is 48 hours at 65 °C and more than 1 hour at 95 °C. Ampligase has been shown to be active for at least 500 thermal cycles (denature 94 °C/anneal, extend 80 °C) or 16 hours of cycling. This exceptional thermostability permits extremely high hybridization stringency and ligation specificity.

Ampligase catalyzes NAD-dependent ligation of adjacent 3´-hydroxylated and 5´-phosphorylated DNA ends. Ampligase has no detectable activity in ligating blunt-ended DNA and has no activity on RNA or RNA:DNA hybrids.

Applications Include:
Gibson Assembly® cloning methods
SNP detection by LDR (Ligase Detection Reaction) or LCR (Ligase Chain Reaction)
Sequence detection or capture by Molecular Inversion Probe (MIP)
Repeat Expansion Detection (RED)
Other ligation-based detection methods
Simultaneous mutagenesis of multiple sites

This enzyme is selective for DNA "nicks", meaning it joins DNA ends that are annealed adjacent to each other on a complementary DNA sequence. Ampligase ideal for applications that require sealing of nicked DNA at elevated temperatures, including Gibson Assembly® and multiple SNP detection methods such as Ligase Chain Reaction (LCR).