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

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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… Click to show full 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 104-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.

Keywords: single cell; scrna seq; cell capture; analysis; capture rna; cell

Journal Title: Methods in molecular biology
Year Published: 2020

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