Single-Cell Multiplex Gene Detection and Sequencing with Microfluidically Generated Agarose Emulsions

Authors

  • Richard Novak,

    1. Center for Exposure Biology, University of California, Berkeley, 307 Lewis Hall, Berkeley, CA 94720 (USA), Fax: (+1) 510-642-3599
    2. UC Berkeley/UC San Francisco Joint Graduate Group in Bioengineering, University of California, Berkeley (USA)
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    • These authors contributed equally.

  • Dr. Yong Zeng,

    1. Center for Exposure Biology, University of California, Berkeley, 307 Lewis Hall, Berkeley, CA 94720 (USA), Fax: (+1) 510-642-3599
    2. Department of Chemistry, University of California, Berkeley (USA)
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    • These authors contributed equally.

  • Dr. Joe Shuga,

    1. Center for Exposure Biology, University of California, Berkeley, 307 Lewis Hall, Berkeley, CA 94720 (USA), Fax: (+1) 510-642-3599
    2. School of Public Health, University of California, Berkeley (USA)
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  • Gautham Venugopalan,

    1. UC Berkeley/UC San Francisco Joint Graduate Group in Bioengineering, University of California, Berkeley (USA)
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  • Prof. Daniel A. Fletcher,

    1. UC Berkeley/UC San Francisco Joint Graduate Group in Bioengineering, University of California, Berkeley (USA)
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  • Prof. Martyn T. Smith,

    1. Center for Exposure Biology, University of California, Berkeley, 307 Lewis Hall, Berkeley, CA 94720 (USA), Fax: (+1) 510-642-3599
    2. School of Public Health, University of California, Berkeley (USA)
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  • Prof. Richard A. Mathies

    Corresponding author
    1. Center for Exposure Biology, University of California, Berkeley, 307 Lewis Hall, Berkeley, CA 94720 (USA), Fax: (+1) 510-642-3599
    2. UC Berkeley/UC San Francisco Joint Graduate Group in Bioengineering, University of California, Berkeley (USA)
    3. Department of Chemistry, University of California, Berkeley (USA)
    • Center for Exposure Biology, University of California, Berkeley, 307 Lewis Hall, Berkeley, CA 94720 (USA), Fax: (+1) 510-642-3599
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  • This research was supported by the trans-NIH Genes, Environment and Health Initiative, Biological Response Indicators of Environmental Systems Center Grant U54 ES016115-01 to M.T.S. and R.A.M., and by Superfund Basic Research Program NIEHS Grant P42 ES004705 to M.T.S. R.N. is supported by an NSF Graduate Research Fellowship, and J.S. is supported by the Canary Foundation and ACS Early Detection Postdoctoral Fellowship.

Abstract

original image

Solitary confinement: The described approach enabled high-throughput DNA preparation followed by multiplex emulsion PCR for the detection and sequencing of genetic targets (see picture). No droplet sorting was required, and single-cell segregation was maintained throughout owing to the incorporation of primer-functionalized beads as amplicon substrates.

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