使用3'端扩增（TPEA）PCR对单细胞进行表达谱分析。
Expression profiling of single cells using 3 prime end amplification (TPEA) PCR.
作者信息
Dixon A K, Richardson P J, Lee K, Carter N P, Freeman T C
机构信息
The Sanger Centre, The Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK,Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QJ, UK.
出版信息
Nucleic Acids Res. 1998 Oct 1;26(19):4426-31. doi: 10.1093/nar/26.19.4426.
The ability to relate the physiological status of individual cells to the complement of genes they express is limited by current methodological approaches for performing these analyses. We report here the development of a robust and reproducible method for amplifying 3' sequences of mRNA derived from single cells and demonstrate that the amplified cDNA, derived from individual human lymphoblastoma cells, can be used for the expression profiling of up to 40 different genes per cell. In addition, we show that 3 prime end amplification (TPEA) PCR can be used to enable the detection of both high and low abundance mRNA species in samples harvested from live neurons in rat brain slices. This procedure will facilitate the study of complex tissue function at the cellular level.
将单个细胞的生理状态与其所表达基因的互补情况联系起来的能力，受到当前用于进行这些分析的方法学途径的限制。我们在此报告一种用于扩增源自单个细胞的mRNA的3'序列的稳健且可重复的方法的开发，并证明源自单个人类淋巴瘤细胞的扩增cDNA可用于每个细胞多达40种不同基因的表达谱分析。此外，我们表明3'末端扩增（TPEA）PCR可用于在从大鼠脑切片中的活神经元收获的样本中检测高丰度和低丰度的mRNA种类。该程序将有助于在细胞水平上研究复杂组织的功能。
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