Fluidigm Corporation, 7000 Shoreline Court, Suite 100, South San Francisco, CA 94080, USA.
Methods. 2013 Jan;59(1):71-9. doi: 10.1016/j.ymeth.2012.10.004. Epub 2012 Oct 16.
The stochastic nature of generating eukaryotic transcripts challenges conventional methods for obtaining and analyzing single-cell gene expression data. In order to address the inherent noise, detailed methods are described on how to collect data on multiple genes in a large number of single cells using microfluidic arrays. As part of a study exploring the effect of genotype on Wnt pathway activation, data were collected for 96 qPCR assays on 1440 lymphoblastoid cells. The description of methods includes preliminary data processing steps. The methods used in the collection and analysis of single-cell qPCR data are contrasted with those used in conventional qPCR.
真核转录本的随机性给获取和分析单细胞基因表达数据的传统方法带来了挑战。为了解决固有噪声的问题，详细描述了如何使用微流控阵列在大量单细胞中收集多个基因的数据。作为一项探索基因型对 Wnt 信号通路激活影响的研究的一部分，对 1440 个淋巴母细胞系进行了 96 个 qPCR 检测。方法描述包括初步的数据处理步骤。对比了单细胞 qPCR 数据的收集和分析方法与传统 qPCR 方法的异同。
