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

Jinhong Kim; Paola A Marignani

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

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

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

Authors

Jinhong Kim¹, Paola A Marignani^{2

3}

Affiliations

¹ Department of Biochemistry and Molecular Biology, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada.
² Department of Biochemistry and Molecular Biology, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada. paola.marignani@dal.ca.
³ Department of Pathology, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada. paola.marignani@dal.ca.

PMID: 35737246
DOI: 10.1007/978-1-0716-2376-3_19

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.

Keywords: Differential gene expression; Fluidigm C1 systems; Protein expression analysis; Validation; qRT-PCR; scRNA-seq.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA, Complementary / genetics
Gene Expression Profiling / methods
High-Throughput Nucleotide Sequencing / methods
RNA / genetics
Sequence Analysis, RNA / methods
Single-Cell Analysis* / methods
Transcriptome*

Substances

DNA, Complementary
RNA