10.1016/j.xpro.2021.100451
https://zenodo.org/records/5846117
oai:zenodo.org:5846117
Dell'Orso, Stefania
Stefania
Dell'Orso
Genomic Technology Section, NIAMS, NIH, Bethesda, MD 20892, USA
Juan, Aster H.
Aster H.
Juan
Laboratory of Muscle Stem Cells and Gene Regulation, NIAMS, NIH, Bethesda, MD 20892, USA
Moiseeva, Victoria
Victoria
Moiseeva
Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), CIBER on Neurodegenerative Diseases (CIBERNED) and ICREA, Barcelona, Spain - Spanish National Center on Cardiovascular Research (CNIC), Madrid, Spain
Garcia-Prat, Laura
Laura
Garcia-Prat
Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), CIBER on Neurodegenerative Diseases (CIBERNED) and ICREA, Barcelona, Spain - Spanish National Center on Cardiovascular Research (CNIC), Madrid, Spain - Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada
Munoz-Canoves, Pura
Pura
Munoz-Canoves
Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), CIBER on Neurodegenerative Diseases (CIBERNED) and ICREA, Barcelona, Spain - Spanish National Center on Cardiovascular Research (CNIC), Madrid, Spain
Sartorelli, Vittorio
Vittorio
Sartorelli
Laboratory of Muscle Stem Cells and Gene Regulation, NIAMS, NIH, Bethesda, MD 20892, USA
Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells
Zenodo
2021
2021-04-15
https://zenodo.org/communities/upgrade-h2020-project
Creative Commons Attribution 4.0 International
Summary
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).
European Commission
10.13039/501100000780
825825
Unlocking Precision Gene Therapy
European Commission
10.13039/501100000780
741966
Tissue regeneration and aging: the decisive quiescent stem-cell state