高通量单细胞实时定量PCR分析

High-Throughput Single-Cell Real-Time Quantitative PCR Analysis.

作者信息

Haim-Vilmovsky Liora

机构信息

EMBL-European Bioinformatics Institute, Wellcome Trust Genome Campus, Cambridge, UK.

Wellcome Sanger Institute, Wellcome Genome Campus, Cambridge, UK.

出版信息

Methods Mol Biol. 2019;1979:177-183. doi: 10.1007/978-1-4939-9240-9_11.

DOI:10.1007/978-1-4939-9240-9_11

PMID:31028638

Abstract

Examining transcriptomics of populations at the single-cell level allows for higher resolution when studying functionality in development, differentiation, and physiology. Real-time quantitative PCR (qPCR) enables a sensitive detection of specific gene expression; however, processing a large number of samples for single-cell research involves a time-consuming process and high reagent costs. Here we describe a protocol for single-cell qPCR using nanofluidic chips. This method reduces the number of handling steps and volumes per reaction, allowing for more samples and genes to be measured.

摘要

在单细胞水平上检查群体的转录组学，在研究发育、分化和生理学中的功能时能够实现更高的分辨率。实时定量PCR（qPCR）能够灵敏地检测特定基因的表达；然而，为单细胞研究处理大量样本涉及耗时的过程和高昂的试剂成本。在此，我们描述了一种使用纳米流体芯片进行单细胞qPCR的方案。该方法减少了每个反应的操作步骤数量和体积，从而能够测量更多的样本和基因。