Cambio Ampligase - Thermostable DNA Ligase

Ampligase® Thermostable DNA Ligase catalyses NAD-dependent ligation of adjacent 3`-hydroxylated and 5`-phosphorylated termini in duplex DNA structures that are stable at high temperatures. Derived from a thermophilic bacterium, the enzyme is stable and active at much higher temperatures than conventional DNA ligases. Its half-life is 48 hours at 65°C and greater than 1hour at 95°C.

Ampligase® DNA Ligase has been shown to be active for at least 500 thermal cycles (94°C/80°C) or 16 hours of cycling. This exceptional thermostability permits extremely high hybridisation stringency and ligation specificity. Ampligase® DNA Ligase has no detectable activity in ligating blunt-ended DNA and has no activity on RNA or RNA:DNA hybrids.

Unit Definition:

One unit of Ampligase® DNA Ligase catalyses the ligation of 50% of the cos sites in 1µg of lambda DNA in 1 minute at 45°C in 1X Ampligase® Reaction Buffer.

Note:

One unit of Ampligase® DNA Ligase is equivalent to at least 15 of the `cohesive end units` or `nick ligation units` defined elsewhere.

Storage Buffer:

50% glycerol containing 50mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1mM EDTA, 1mM DTT, and 0.1% Triton® X-100.

Ampligase® 10X Reaction Buffer:

200 mM Tris-HCl (pH 8.3), 250mM KCl, 100mM MgCl2, 5mM NAD, and 0.1% Triton® X-100.

Quality Control:

Ampligase® Thermostable DNA Ligase is assayed for the absence of blunt-end ligation activity by ligating SmalI-digested bacteriophage lambda DNA at 45°C; ligation occurs only at cos sites as determined by gel electrophoresis. Ampligase® DNA Ligase is free of detectable exo- and endonuclease and RNase activities.