Microfluidic Single-Cell mRNA Isolation and Analysis

Single-cell gene expression analysis holds great promise for studying diverse biological systems, but methodology to process these precious samples in a reproducible, quantitative, and parallel fashion remains challenging. Here, we utilize microfluidics to isolate picogram and subpicogram mRNA templ...

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Veröffentlicht in:Analytical chemistry (Washington) 2006-05, Vol.78 (9), p.3084-3089
Hauptverfasser: Marcus, Joshua S., Anderson, W. French, Quake, Stephen R.
Format: Artikel
Sprache:eng
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Zusammenfassung:Single-cell gene expression analysis holds great promise for studying diverse biological systems, but methodology to process these precious samples in a reproducible, quantitative, and parallel fashion remains challenging. Here, we utilize microfluidics to isolate picogram and subpicogram mRNA templates, as well as to synthesize cDNA from these templates. We demonstrate single-cell mRNA isolation and cDNA synthesis, provide quantitative calibrations for each step in the process, and measure gene expression in individual cells. The techniques presented here form the foundation for highly parallel single-cell gene expression studies.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac0519460