Deciphering the Early Mouse Embryo Transcriptome by Low-Input RNA-Seq.
Fiche publication
Date publication
janvier 2021
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Lien Pubmed
Résumé
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.
Mots clés
Embryonic gene expression, Low-input transcriptomics, Single-cell RNA-seq, Single-embryo RNA-seq, Spike-In, scRNA-seq
Référence
Methods Mol Biol. 2021 ;2214:189-205