西亚试剂：High-throughput microfluidic single-cell RT-qPCR

High-throughput microfluidic single-cell RT-qPCR

White, Adam K.; VanInsberghe, Michael; Petriv, Oleh I.; Hamidi, Mani; Sikorski, Darek; Marra, Marco A.; Piret, James; Aparicio, Samuel; Hansen, Carl L.

A long-sought milestone in microfluidics research has been the development of integrated technology for scalable analysisof transcription in single cells. Here we present a fully integrated microfluidic device capable of performing high-precisionRT-qPCR measurements of gene expression from hundreds of single cells per run. Our device executes all steps of single-cellprocessing, including cell capture, cell lysis, reverse transcription, and quantitative PCR. In addition to higher throughputand reduced cost, we show that nanoliter volume processing reduced measurement noise, increased sensitivity, and providedsingle nucleotide specificity. We apply this technology to 3,300 single-cell measurements of (i) miRNA expression in K562 cells, (ii) coregulation of a miRNA and one of its target transcripts during differentiation in embryonic stem cells, and (iii) single nucleotide variant detection in primary lobular breast cancer cells. The core functionality established here providesthe foundation from which a variety of on-chip single-cell transcription analyses will be developed.

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