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Single-Cell Isolation from Regenerating Murine Muscles for RNA-Sequencing Analysis.

ABSTRACT: Single-cell RNA sequencing (scRNA-seq) is a powerful technique for deconvoluting and clustering thousands of otherwise intermingled cells based on their gene expression. Here, we present a complete protocol for the unbiased evaluation of regenerating murine skeletal muscle using scRNA-seq. The skeletal muscle is unique in its cellular composition as being primarily multinucleated muscle cells (myofibers). This protocol focuses on isolating mononuclear cells from muscle for subsequent scRNA-seq analysis and can be modified to assess cell populations in other tissues of interest. For complete details on the use and execution of this protocol, please refer to Liu et al. (2015) and Oprescu et al. (2020).

SUBMITTER: Oprescu SN

PROVIDER: S-EPMC7580090 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Single-Cell Isolation from Regenerating Murine Muscles for RNA-Sequencing Analysis.

Oprescu Stephanie N SN Yue Feng F Kuang Shihuan S

STAR protocols 20200612 2

Single-cell RNA sequencing (scRNA-seq) is a powerful technique for deconvoluting and clustering thousands of otherwise intermingled cells based on their gene expression. Here, we present a complete protocol for the unbiased evaluation of regenerating murine skeletal muscle using scRNA-seq. The skeletal muscle is unique in its cellular composition as being primarily multinucleated muscle cells (myofibers). This protocol focuses on isolating mononuclear cells from muscle for subsequent scRNA-seq a ...[more]

PMID: 33111097

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