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

Membres identifiés du Cancéropôle Est :
Pr FAUQUE Patricia


Tous les auteurs :
Pérez-Palacios R, Fauque P, Teissandier A, Bourc'his D

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