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: Dell'Orso, Stefania, Juan, Aster H., Moiseeva, Victoria, 1992-, García Prat, Laura, Muñoz Cánoves, Pura, 1962-, Sartorelli, Vittorio
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/47411
Acceso en línea:http://hdl.handle.net/10230/47411
http://dx.doi.org/10.1016/j.xpro.2021.100451
Access Level:acceso abierto
Palabra clave:Cell differentiation
Cell isolation
Flow Cytometry/Mass Cytometry
Molecular biology
RNA-seq
Stem cells
Descripción
Sumario: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).