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 1 hour 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. ¹⁰ This exceptional thermostability permits extremely high hybridization 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.

Applications

• Ligation amplification (Ligase Chain Reaction, LCR) ^1-7
• Repeat Expansion Detection (RED) ⁷
• Simultaneous mutagenesis of multiple sites ^8,9
• Other ligation-based detection methods

References

1. Landegren, U. et al. (1988) Science 242, 229.
2. Wu, D.Y. and Wallace, R.B. (1989) Genomics 4, 560.
3. Barany, F. (1991) Proc. Natl. Acad. Sci. USA 88, 189.
4. Birkenmeyer, L. and Armstrong, A.S. (1992) J. Clin. Micro. 30, 3089.
5. Dille, B.J. et al. (1993) J. Clin. Micro. 31, 729
6. Nakazawa, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 360.
7. Sirugo, G. and Kidd, K. (1995) Epicentre Forum 2(3), 1.
8. Moore, D. and Michael, S. (1995) Epicentre Forum 2(4), 4.
9. Rouwendal, G.J. et al. (1993) BioTechniques 15, 68.
10. Schalling, M. et al. (1993) Nature Genetics 4, 135.
11. Hardenbol, P. et al. (2003) Nature Biotechnology 21, 673.