Real-Time Nucleic Acid Sequence-Based Amplification in Nanoliter Volumes

  • Gulliksen A
  • Solli L
  • Karlsen F
 et al. 
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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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Authors

  • Anja Gulliksen

  • Lars Solli

  • Frank Karlsen

  • Henrik Rogne

  • Eivind Hovig

  • Trine Nordstrøm

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