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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.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.issn 2666-1667
dc.identifier.uri http://hdl.handle.net/10230/47411
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.language.iso eng
dc.publisher Elsevier
dc.relation.ispartof STAR Protoc. 2021;2(2):100451
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.uri http://creativecommons.org/licenses/by-nc-nd/4.0/
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.identifier.doi http://dx.doi.org/10.1016/j.xpro.2021.100451
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.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.accessRights info:eu-repo/semantics/openAccess
dc.type.version info:eu-repo/semantics/publishedVersion

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