Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells

It remains challenging to generate reproducible, high-quality cDNA libraries from RNA derived from rare cell populations. Here, we describe a protocol for high-throughput RNA-seq library preparation, including isolation of 200 skeletal muscle stem cells from mouse tibialis anterior muscle by fluores...

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Detalles Bibliográficos
Autores principales: Dell’Orso, Stefania, Juan, Aster H., Moiseeva, Victoria, García-Prat, Laura, Muñoz-Cánoves, Pura, Sartorelli, Vittorio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079445/
https://www.ncbi.nlm.nih.gov/pubmed/33937872
http://dx.doi.org/10.1016/j.xpro.2021.100451
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author Dell’Orso, Stefania
Juan, Aster H.
Moiseeva, Victoria
García-Prat, Laura
Muñoz-Cánoves, Pura
Sartorelli, Vittorio
author_facet Dell’Orso, Stefania
Juan, Aster H.
Moiseeva, Victoria
García-Prat, Laura
Muñoz-Cánoves, Pura
Sartorelli, Vittorio
author_sort Dell’Orso, Stefania
collection PubMed
description It remains challenging to generate reproducible, high-quality cDNA libraries from RNA derived from rare cell populations. Here, we describe a protocol for high-throughput RNA-seq library preparation, including isolation of 200 skeletal muscle stem cells from mouse tibialis anterior muscle by fluorescence-activated cell sorting and cDNA preparation. We also describe RNA extraction and cDNA preparation from differentiating mouse embryonic stem cells. For complete details on the use and execution of this protocol, please refer to Juan et al. (2016) and Garcia-Prat et al. (2016).
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spelling pubmed-80794452021-04-29 Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells Dell’Orso, Stefania Juan, Aster H. Moiseeva, Victoria García-Prat, Laura Muñoz-Cánoves, Pura Sartorelli, Vittorio STAR Protoc Protocol It remains challenging to generate reproducible, high-quality cDNA libraries from RNA derived from rare cell populations. Here, we describe a protocol for high-throughput RNA-seq library preparation, including isolation of 200 skeletal muscle stem cells from mouse tibialis anterior muscle by fluorescence-activated cell sorting and cDNA preparation. We also describe RNA extraction and cDNA preparation from differentiating mouse embryonic stem cells. For complete details on the use and execution of this protocol, please refer to Juan et al. (2016) and Garcia-Prat et al. (2016). Elsevier 2021-04-15 /pmc/articles/PMC8079445/ /pubmed/33937872 http://dx.doi.org/10.1016/j.xpro.2021.100451 Text en 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
Dell’Orso, Stefania
Juan, Aster H.
Moiseeva, Victoria
García-Prat, Laura
Muñoz-Cánoves, Pura
Sartorelli, Vittorio
Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
title Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
title_full Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
title_fullStr Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
title_full_unstemmed Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
title_short Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
title_sort protocol for rna-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079445/
https://www.ncbi.nlm.nih.gov/pubmed/33937872
http://dx.doi.org/10.1016/j.xpro.2021.100451
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