Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
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- dc.contributor.author Dell'Orso, Stefania
- dc.contributor.author Juan, Aster H.
- dc.contributor.author Moiseeva, Victoria, 1992-
- dc.contributor.author García Prat, Laura
- dc.contributor.author Muñoz Cánoves, Pura, 1962-
- dc.contributor.author Sartorelli, Vittorio
- dc.date.accessioned 2021-05-13T06:55:07Z
- dc.date.available 2021-05-13T06:55:07Z
- dc.date.issued 2021
- dc.description.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).
- dc.description.sponsorship Funds to V.S. were provided by the NIAMS-IRP through NIH grants AR041126 and AR041164. Work in P.M.-C’s laboratory was supported by MINECO-RTI2018-096068, ERC-AdG-741966, LaCaixa-HEALTH-HR17-00040, UPGRADE-H2020-825825 and Marató-TV3, MDM-2014-0370, and SEV-2015-0505. Authors acknowledge technical support from the UPF/CRG and CNIC flow cytometry units.
- dc.format.mimetype application/pdf
- dc.identifier.citation 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;2(2):100451. DOI: 10.1016/j.xpro.2021.100451
- dc.identifier.doi http://dx.doi.org/10.1016/j.xpro.2021.100451
- dc.identifier.issn 2666-1667
- dc.identifier.uri http://hdl.handle.net/10230/47411
- dc.language.iso eng
- dc.publisher Elsevier
- dc.relation.ispartof STAR Protoc. 2021;2(2):100451
- dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/741966
- dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/RTI2018-096068
- dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/825825
- dc.rights © This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
- dc.rights.accessRights info:eu-repo/semantics/openAccess
- dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/4.0/
- dc.subject.keyword Cell differentiation
- dc.subject.keyword Cell isolation
- dc.subject.keyword Flow Cytometry/Mass Cytometry
- dc.subject.keyword Molecular biology
- dc.subject.keyword RNA-seq
- dc.subject.keyword Stem cells
- dc.title Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
- dc.type info:eu-repo/semantics/article
- dc.type.version info:eu-repo/semantics/publishedVersion