Marcus Joshua S, Anderson W French, Quake Stephen R
Biochemistry and Molecular Biophysics, California Institute of Technology, MS 128-95, Pasadena, California 91125, USA.
Anal Chem. 2006 May 1;78(9):3084-9. doi: 10.1021/ac0519460.
Single-cell gene expression analysis holds great promise for studying diverse biological systems, but methodology to process these precious samples in a reproducible, quantitative, and parallel fashion remains challenging. Here, we utilize microfluidics to isolate picogram and subpicogram mRNA templates, as well as to synthesize cDNA from these templates. We demonstrate single-cell mRNA isolation and cDNA synthesis, provide quantitative calibrations for each step in the process, and measure gene expression in individual cells. The techniques presented here form the foundation for highly parallel single-cell gene expression studies.
单细胞基因表达分析在研究各种生物系统方面具有巨大潜力，但以可重复、定量和平行的方式处理这些珍贵样本的方法仍然具有挑战性。在这里，我们利用微流控技术来分离皮克和亚皮克级的mRNA模板，并从这些模板合成cDNA。我们展示了单细胞mRNA分离和cDNA合成，为该过程的每个步骤提供了定量校准，并测量了单个细胞中的基因表达。本文介绍的技术为高度平行的单细胞基因表达研究奠定了基础。
