Droplet-Based Microfluidic Systems for High-Throughput Single DNA Molecule Isothermal Amplification and Analysis
mediaposted on 15.06.2009, 00:00 by Linas Mazutis, Ali Fallah Araghi, Oliver J. Miller, Jean-Christophe Baret, Lucas Frenz, Agnes Janoshazi, Valérie Taly, Benjamin J. Miller, J. Brian Hutchison, Darren Link, Andrew D. Griffiths, Michael Ryckelynck
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.