Droplet-based microfluidic systems for high-throughput single DNA molecule isothermal amplification and analysis.

Linas Mazutis 1 Ali Fallah Araghi 1 Oliver Miller 1 Jean-Christophe Baret 1 Lucas Frenz 1 Agnes Janoshazi 1, 2 Valérie Taly 1 Benjamin Miller 3 J Brian Hutchison 3 Darren Link 3 Andrew Griffiths 1, 2 Michael Ryckelynck 1, 2
1 ISIS - Institut de Science et d'ingénierie supramoléculaires
2 IGBMC - Institut de génétique et biologie moléculaire et cellulaire
3 RainDance Technologies, Inc.
Abstract : We have developed a method for high-throughput isothermal amplification of single DNA molecules in a droplet-based microfluidic system. DNA amplification in droplets was analyzed using an intercalating fluorochrome, allowing fast and accurate "digital" quantification of the template DNA based on the Poisson distribution of DNA molecules in droplets. The clonal amplified DNA in each 2 pL droplet was further analyzed by measuring the enzymatic activity of the encoded proteins after fusion with a 15 pL droplet containing an in vitro translation system.
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Journal articles
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https://www.hal.inserm.fr/inserm-00420178
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Submitted on : Monday, September 28, 2009 - 1:29:20 PM
Last modification on : Friday, September 27, 2019 - 1:58:02 PM

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Linas Mazutis, Ali Fallah Araghi, Oliver Miller, Jean-Christophe Baret, Lucas Frenz, et al.. Droplet-based microfluidic systems for high-throughput single DNA molecule isothermal amplification and analysis.. Analytical Chemistry, American Chemical Society, 2009, 81 (12), pp.4813-21. ⟨10.1021/ac900403z⟩. ⟨inserm-00420178⟩

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