Journal
ANALYTICAL CHEMISTRY
Volume 85, Issue 16, Pages 8016-8021Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac402057q
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Funding
- NSF [DBI-1253293]
- California Institute for Quantitative Biosciences [QB3]
- Rogers Family Foundation
- UCSF/Sandler Foundation Program for Breakthrough Biomedical Research
- University of California
- Direct For Biological Sciences
- Div Of Biological Infrastructure [1253293] Funding Source: National Science Foundation
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The behaviors of complex biological systems are often dictated by the properties of their heterogeneous and sometimes rare cellular constituents. Correspondingly, the analysis of individual cells from a heterogeneous population can reveal information not obtainable by ensemble measurements. Reverse-transcriptase polymerase chain reaction (RT-PCR) is a widely used method that enables transcriptional profiling and sequencing analysis on bulk populations of cells. Major barriers to successfully implementing this technique for mammalian single-cell studies are the labor, cost, and low-throughput associated with current approaches. In this report, we describe a novel droplet-based microfluidic system for performing similar to 50000 single-cell RT-PCR reactions in a single experiment while consuming a minimal amount of reagent. Using cell type-specific staining and TaqMan RT-PCR probes, we demonstrate the identification of specific cells from a mixed human cell population. The throughput, robust detection rate and specificity of this method makes it well-suited for characterizing large, heterogeneous populations of cells at the transcriptional level.
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