RNA Sequencing of Single Myofibers from Mus musculus

Darren M Blackburn; Felicia Lazure; Vahab D Soleimani

doi:10.21769/BioProtoc.3525

RNA Sequencing of Single Myofibers from Mus musculus

Bio Protoc. 2020 Feb 20;10(4):e3525. doi: 10.21769/BioProtoc.3525.

Authors

Darren M Blackburn^{1

2}, Felicia Lazure^{1

2}, Vahab D Soleimani^{1

2}

Affiliations

¹ Department of Human Genetics, McGill University, 3640 rue University, Montreal, QC, H3A 0C7 Canada.
² Molecular and Regenerative Medicine Axis, Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 Chemin de la Cote-Sainte-Catherine, Montreal, QC, H3T 1E2, Canada.

Abstract

Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the transcriptome of myofibers we combine single myofiber isolation with SMART-Seq to provide high resolution genome wide expression of a single myofiber.

Keywords: Gene expression analysis; SMART-Seq; Single myofiber; Skeletal muscle; Transcriptomics.