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Dell'Orso S, Juan AH, Moiseeva V, et al. Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells. STAR Protoc. 2021;2(2):100451doi: 10.1016/j.xpro.2021.100451.
Dell'Orso, S., Juan, A. H., Moiseeva, V., García-Prat, L., Muñoz-Cánoves, P., & Sartorelli, V. (2021). Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells. STAR protocols, 2(2), 100451. https://doi.org/10.1016/j.xpro.2021.100451
Dell'Orso, Stefania, et al. "Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells." STAR protocols vol. 2,2 (2021): 100451. doi: https://doi.org/10.1016/j.xpro.2021.100451
Dell'Orso S, Juan AH, Moiseeva V, García-Prat L, Muñoz-Cánoves P, Sartorelli V. Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells. STAR Protoc. 2021 Apr 15;2(2):100451. doi: 10.1016/j.xpro.2021.100451. eCollection 2021 Jun 18. PMID: 33937872; PMCID: PMC8079445.
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STAR Protoc. 2021 Apr 15;2(2):100451. doi: 10.1016/j.xpro.2021.100451. eCollection 2021 Jun 18.

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

STAR protocols

Stefania Dell'Orso, Aster H Juan, Victoria Moiseeva, Laura García-Prat, Pura Muñoz-Cánoves, Vittorio Sartorelli

Affiliations

  1. Genomic Technology Section, NIAMS, NIH, Bethesda, MD 20892, USA.
  2. Laboratory of Muscle Stem Cells and Gene Regulation, NIAMS, NIH, Bethesda, MD 20892, USA.
  3. Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), CIBER on Neurodegenerative Diseases (CIBERNED) and ICREA, Barcelona, Spain.
  4. Spanish National Center on Cardiovascular Research (CNIC), Madrid, Spain.
  5. Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada.

PMID: 33937872 PMCID: PMC8079445 DOI: 10.1016/j.xpro.2021.100451

Abstract

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).

Keywords: Cell Differentiation; Cell isolation; Flow Cytometry/Mass Cytometry; Molecular Biology; RNA-seq; Stem Cells

Conflict of interest statement

The authors declare no competing interests.

References

  1. Nature. 2014 Feb 20;506(7488):316-21 - PubMed
  2. Nature. 2016 Jan 7;529(7584):37-42 - PubMed
  3. Cell Rep. 2016 Oct 25;17(5):1369-1382 - PubMed
  4. Nat Cell Biol. 2020 Nov;22(11):1307-1318 - PubMed

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