Abstract
Early preimplantation embryos are precious and scarce samples that contain limited numbers of cells, which can be problematic for quantitative gene expression analyses. Nonetheless, low-input genome-wide techniques coupled with cDNA amplification steps have become a gold standard for RNA profiling of as minimal as a single blastomere. Here, we describe a single-cell/single-embryo RNA sequencing (RNA-seq) method, from embryo collection to sample validation steps prior to DNA library preparation and sequencing. Key quality controls and external Spike-In normalization approaches are also detailed.
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Acknowledgments
The laboratory of D.B. is part of the Laboratoires d’Excellence (LABEX) entitled DEEP (11-LBX0044). P.F’s research is supported by Agence Nationale pour la Recherche (ANR-17-CE12-0014).
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Pérez-Palacios, R., Fauque, P., Teissandier, A., Bourc’his, D. (2021). Deciphering the Early Mouse Embryo Transcriptome by Low-Input RNA-Seq. In: Ancelin, K., Borensztein, M. (eds) Epigenetic Reprogramming During Mouse Embryogenesis. Methods in Molecular Biology, vol 2214. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0958-3_13
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DOI: https://doi.org/10.1007/978-1-0716-0958-3_13
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0957-6
Online ISBN: 978-1-0716-0958-3
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