Single-Cell Capture, RNA-seq, and Transcriptome Analysis from the Neural Retina.

 
TitleSingle-Cell Capture, RNA-seq, and Transcriptome Analysis from the Neural Retina.
Publication TypeJournal Article
Year of Publication2020
AuthorsDharmat, R, Kim, S, Li, Y, Chen, R
JournalMethods Mol Biol
Volume2092
Pagination159-186
Date Published2020
ISSN1940-6029
Abstract

Single-cell RNA sequencing (scRNA-seq) is an emerging technology that can address the challenge of cellular heterogeneity. In the last decade, the cost per cell has been dramatically reduced, and the throughput has been increased by 10-fold. Like many other tissues, the retina is highly heterogeneous with an estimated of over 100 subtypes of neuronal cells. Here, we describe the current techniques to perform scRNA-seq on the adult retinal tissue including retinal dissection, retinal dissociation, assessment of cell population, cDNA synthesis, library construction, and next-generation sequencing. In addition, we introduce a workflow of scRNA-seq data analysis using open-source tools.

DOI10.1007/978-1-0716-0175-4_12
Alternate JournalMethods Mol. Biol.
PubMed ID31786788