Real-time nucleic acid sequence-based amplification in nanoliter volumes |
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Authors: | Gulliksen Anja Solli Lars Karlsen Frank Rogne Henrik Hovig Eivind Nordstrøm Trine Sirevåg Reidun |
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Affiliation: | NorChip AS, Industriveien 8, 3490 Klokkarstua, Norway. Anja.Gulliksen@norchip.com |
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Abstract: | Real-time nucleic acid sequence-based amplification (NASBA) is an isothermal method specifically designed for amplification of RNA. Fluorescent molecular beacon probes enable real-time monitoring of the amplification process. Successful identification, utilizing the real-time NASBA technology, was performed on a microchip with oligonucleotides at a concentration of 1.0 and 0.1 microM, in 10- and 50-nL reaction chambers, respectively. The microchip was developed in a silicon-glass structure. An instrument providing thermal control and an optical detection system was built for amplification readout. Experimental results demonstrate distinct amplification processes. Miniaturized real-time NASBA in microchips makes high-throughput diagnostics of bacteria, viruses, and cancer markers possible, at reduced cost and without contamination. |
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