Extraction and sequencing of single nuclei from murine skeletal muscles

Single-nucleus RNA sequencing allows the profiling of gene expression in isolated nuclei. Here, we describe a step-by-step protocol optimized for adult mouse skeletal muscles. This protocol provides two main advantages compared to the widely used single-cell protocol. First, it allows us to sequence...

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Autores principales: Santos, Matthieu Dos, Gioftsidi, Stamatia, Backer, Stéphanie, Machado, Léo, Relaix, Frederic, Maire, Pascal, Mourikis, Philippos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339382/
https://www.ncbi.nlm.nih.gov/pubmed/34382019
http://dx.doi.org/10.1016/j.xpro.2021.100694
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author Santos, Matthieu Dos
Gioftsidi, Stamatia
Backer, Stéphanie
Machado, Léo
Relaix, Frederic
Maire, Pascal
Mourikis, Philippos
author_facet Santos, Matthieu Dos
Gioftsidi, Stamatia
Backer, Stéphanie
Machado, Léo
Relaix, Frederic
Maire, Pascal
Mourikis, Philippos
author_sort Santos, Matthieu Dos
collection PubMed
description Single-nucleus RNA sequencing allows the profiling of gene expression in isolated nuclei. Here, we describe a step-by-step protocol optimized for adult mouse skeletal muscles. This protocol provides two main advantages compared to the widely used single-cell protocol. First, it allows us to sequence the myonuclei of the multinucleated myofibers. Second, it circumvents the cell-dissociation-induced transcriptional modifications. For complete details on the use and execution of this protocol, please refer to Dos Santos et al. (2020) and Machado, Geara et al. (2021).
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spelling pubmed-83393822021-08-10 Extraction and sequencing of single nuclei from murine skeletal muscles Santos, Matthieu Dos Gioftsidi, Stamatia Backer, Stéphanie Machado, Léo Relaix, Frederic Maire, Pascal Mourikis, Philippos STAR Protoc Protocol Single-nucleus RNA sequencing allows the profiling of gene expression in isolated nuclei. Here, we describe a step-by-step protocol optimized for adult mouse skeletal muscles. This protocol provides two main advantages compared to the widely used single-cell protocol. First, it allows us to sequence the myonuclei of the multinucleated myofibers. Second, it circumvents the cell-dissociation-induced transcriptional modifications. For complete details on the use and execution of this protocol, please refer to Dos Santos et al. (2020) and Machado, Geara et al. (2021). Elsevier 2021-07-28 /pmc/articles/PMC8339382/ /pubmed/34382019 http://dx.doi.org/10.1016/j.xpro.2021.100694 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Santos, Matthieu Dos
Gioftsidi, Stamatia
Backer, Stéphanie
Machado, Léo
Relaix, Frederic
Maire, Pascal
Mourikis, Philippos
Extraction and sequencing of single nuclei from murine skeletal muscles
title Extraction and sequencing of single nuclei from murine skeletal muscles
title_full Extraction and sequencing of single nuclei from murine skeletal muscles
title_fullStr Extraction and sequencing of single nuclei from murine skeletal muscles
title_full_unstemmed Extraction and sequencing of single nuclei from murine skeletal muscles
title_short Extraction and sequencing of single nuclei from murine skeletal muscles
title_sort extraction and sequencing of single nuclei from murine skeletal muscles
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339382/
https://www.ncbi.nlm.nih.gov/pubmed/34382019
http://dx.doi.org/10.1016/j.xpro.2021.100694
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