Extraction and sequencing of single nuclei from murine skeletal muscles
Summary: 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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721004019 |
| _version_ | 1819082199510548480 |
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| author | Matthieu Dos Santos Stamatia Gioftsidi Stéphanie Backer Léo Machado Frederic Relaix Pascal Maire Philippos Mourikis |
| author_facet | Matthieu Dos Santos Stamatia Gioftsidi Stéphanie Backer Léo Machado Frederic Relaix Pascal Maire Philippos Mourikis |
| author_sort | Matthieu Dos Santos |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-21T20:12:53Z |
| format | Article |
| id | doaj.art-012167ae4a2b4b66a5f210b57136fc20 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-21T20:12:53Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-012167ae4a2b4b66a5f210b57136fc202022-12-21T18:51:41ZengElsevierSTAR Protocols2666-16672021-09-0123100694Extraction and sequencing of single nuclei from murine skeletal musclesMatthieu Dos Santos0Stamatia Gioftsidi1Stéphanie Backer2Léo Machado3Frederic Relaix4Pascal Maire5Philippos Mourikis6Université de Paris, Institut Cochin, INSERM, CNRS, 75014 Paris, France; Corresponding authorUniv Paris Est Créteil, INSERM, IMRB, 94010 Créteil, FranceUniversité de Paris, Institut Cochin, INSERM, CNRS, 75014 Paris, FranceUniv Paris Est Créteil, INSERM, IMRB, 94010 Créteil, FranceUniv Paris Est Créteil, INSERM, IMRB, 94010 Créteil, France; EnvA, IMRB, 94700 Maisons-Alfort, France; EFS, IMRB, 94010 Creteil, France; AP-HP, Hopital Mondor, Service d’histologie, 94010 Creteil, FranceUniversité de Paris, Institut Cochin, INSERM, CNRS, 75014 Paris, France; Corresponding authorUniv Paris Est Créteil, INSERM, IMRB, 94010 Créteil, France; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166721004019Cell isolationSingle CellFlow Cytometry/Mass CytometryRNAseqStem CellsCell Differentiation |
| spellingShingle | Matthieu Dos Santos Stamatia Gioftsidi Stéphanie Backer Léo Machado Frederic Relaix Pascal Maire Philippos Mourikis Extraction and sequencing of single nuclei from murine skeletal muscles STAR Protocols Cell isolation Single Cell Flow Cytometry/Mass Cytometry RNAseq Stem Cells Cell Differentiation |
| 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 | Cell isolation Single Cell Flow Cytometry/Mass Cytometry RNAseq Stem Cells Cell Differentiation |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721004019 |
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