title: Droplet-based microfluidic systems for high-throughput single DNA molecule isothermal amplification and analysis.
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.
authors.affiliation: Institut de Science et d'Ingenierie Supramoleculaire, Universite de Strasbourg, CNRS UMR 7006, 8 allee Gaspard Monge, 67083 Strasbourg Cedex, France.
authors.forename: Linas
authors.initials: L
authors.lastname: Mazutis
authors.forename: Ali Fallah
authors.initials: AF
authors.lastname: Araghi
authors.forename: Oliver J
authors.initials: OJ
authors.lastname: Miller
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Baret
authors.forename: Lucas
authors.initials: L
authors.lastname: Frenz
authors.forename: Agnes
authors.initials: A
authors.lastname: Janoshazi
authors.forename: Valérie
authors.initials: V
authors.lastname: Taly
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Miller
authors.forename: J Brian
authors.initials: JB
authors.lastname: Hutchison
authors.forename: Darren
authors.initials: D
authors.lastname: Link
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Griffiths
authors.forename: Michael
authors.initials: M
authors.lastname: Ryckelynck
mesh_terms: D004247:DNA / Q000032:analysis*; D007364:Intercalating Agents / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D013696:Temperature
chemical_list: D007364:Intercalating Agents; D004247:DNA