微流控单细胞实时 PCR 用于比较基因表达模式的分析。
Microfluidic single-cell real-time PCR for comparative analysis of gene expression patterns.
机构信息
Department of Medicine, Stanford University School of Medicine, California, USA.
出版信息
Nat Protoc. 2012 Apr 5;7(5):829-38. doi: 10.1038/nprot.2012.021.
Abstract
Single-cell quantitative real-time PCR (qRT-PCR) combined with high-throughput arrays allows the analysis of gene expression profiles at a molecular level in approximately 11 h after cell sample collection. We present here a high-content microfluidic real-time platform as a powerful tool for comparatively investigating the regulation of developmental processes in single cells. This approach overcomes the limitations involving heterogeneous cell populations and sample amounts, and may shed light on differential regulation of gene expression in normal versus disease-related contexts. Furthermore, high-throughput single-cell qRT-PCR provides a standardized, comparative assay for in-depth analysis of the mechanisms underlying human pluripotent stem cell self-renewal and differentiation.
摘要
单细胞实时定量 PCR(qRT-PCR) 结合高通量芯片可在细胞样本采集后约 11 小时内在分子水平上分析基因表达谱。我们在此介绍一种高内涵微流控实时平台，它是一种强大的工具，可用于比较研究单细胞中发育过程的调控。这种方法克服了涉及异质细胞群体和样本量的限制，并可能揭示正常和疾病相关情况下基因表达差异调控的机制。此外，高通量单细胞 qRT-PCR 为深入分析人类多能干细胞自我更新和分化的机制提供了一种标准化、可比的检测方法。
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