Optical mapping of single-molecule human DNA in disposable, mass-produced all-polymer

Peter Friis Østergaard, Joanna Lopacinska-Jørgensen, Jonas Nyvold Pedersen, Niels Tommerup, Anders Kristensen, Henrik Flyvbjerg, Asli Silahtaroglu, Rodolphe Marie, Rafael J. Taboryski

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    Abstract

    We demonstrate all-polymer injection molded devices for optical mapping of denaturation–renaturation (DR) patterns on long, single DNA-molecules from the human genome. The devices have channels with ultra-low aspect ratio, only 110 nm deep while 20 μm wide, and are superior to the silica devices used previously in the field. With these polymer devices, we demonstrate on-chip recording of DR images of DNA-molecules stretched to more than 95% of their contour length. The stretching is done by opposing flows Marie et al (2013 Proc. Natl Acad. Sci. USA 110 4893–8). The performance is validated by mapping 20 out of 24 Mbp-long DNA fragments to the human reference genome. We optimized fabrication of the devices to a yield exceeding 95%. This permits a substantial economies-of-scale driven cost-reduction, leading to device costs as low as 3 USD per device, about a factor 70 lower than the cost of silica devices. This lowers the barrier to a wide use of DR mapping of native, megabase-size DNA molecules, which has a huge potential as a complementary method to next-generation sequencing.
    Original languageEnglish
    JournalJournal of Micromechanics and Microengineering
    Volume25
    Pages (from-to)105002
    Number of pages8
    ISSN0960-1317
    DOIs
    Publication statusPublished - 2015

    Keywords

    • Injection molding
    • Human DNA mapping
    • Lab-on-a-chip

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