使用Fluidigm C1平台进行单细胞RNA测序分析以表征异质转录组

Single-Cell RNA Sequencing Analysis Using Fluidigm C1 Platform for Characterization of Heterogeneous Transcriptomes.

作者信息

Kim Jinhong, Marignani Paola A

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada.

Department of Pathology, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada.

出版信息

Methods Mol Biol. 2022;2508:261-278. doi: 10.1007/978-1-0716-2376-3_19.

DOI:10.1007/978-1-0716-2376-3_19

PMID:35737246

Abstract

Single-cell RNA sequencing (scRNA-seq) is an innovative technology that can be used to characterize transcriptome heterogeneity at the single-cell resolution. The Fluidigm C1 is an automated system for preparing complementary deoxyribonucleic acid (cDNA) samples that have been synthesized and preamplified from single cells for next-generation sequencing (NGS) analysis. Herein, we detail a workflow and protocol for scRNA-seq analysis of 3'-end enriched cDNA libraries and introduce strategies for validation of differential gene expression.

摘要

单细胞RNA测序（scRNA-seq）是一项创新技术，可用于在单细胞分辨率下表征转录组异质性。Fluidigm C1是一种自动化系统，用于制备已从单细胞合成并预扩增的互补脱氧核糖核酸（cDNA）样本，以进行下一代测序（NGS）分析。在此，我们详细介绍了3'端富集cDNA文库的scRNA-seq分析工作流程和方案，并介绍了差异基因表达验证策略。