High-throughput single-cell PCR using microfluidic emulsions

POSTER

Abstract

The human gut and other environmental samples contain large populations of diverse bacteria that are poorly characterized and unculturable, yet have many functions relevant to human health. Our goal is to identify exactly which species carry some gene of interest, such as a carbohydrate metabolism gene. Conventional metagenomic assays sequence DNA extracted in bulk from populations of mixed cell types, and are therefore unable to associate a gene of interest with a species-identifying 16S gene, to determine that the two genes originated from the same cell. We solve this problem by microfluidically encapsulating single bacteria cells in drops, using PCR to amplify the two genes inside any drop whose encapsulated cell contains both genes, and sequencing the DNA from those drops that contain both amplification products.

*Supported by NSF and Harvard MRSEC.

Authors

  • Mira Guo

    • Harvard University

  • Linas Mazutis

    • Harvard University

  • Jeremy Agresti

    • Harvard University

  • Morten Sommer

    • Harvard University

  • Gautam Dantas

    • Harvard University

  • George Church

    • Harvard University

  • Peter Turnbaugh

    • Harvard University

  • David Weitz

    • Harvard University
    • Department of Physics and Division of Engineering and Applied Science, Harvard University, Cambridge, MA 02138
    • School of Engineering and Applied Sciences, Harvard University
    • Department of Physics, Harvard University, Cambridge, MA, United States
    • Department of Physics and School of Engineering and Applied Sciences, Harvard University, Cambridge, MA