GSM6543170: Placode1; Mus musculus; RNA-Seq
Epithelium and mesenchyme of dorsal skins were enzymatically separated using Dispase II (Roche, Tokyo, Japan). Epithelial sheets of embryos from the same litter were pooled (3-4 embryos per sample) and dissociated to single cell suspension by TrypLE Select Enzyme (Life Technologies/Gibco). Cell were stained with rat anti-MAdCAM-1 (BD Pharmingen), followed by anti-rat Alexa Fluor 488 - conjugated antibody (Life Technologies). Live total epithelium cells or placode-enriched (MAdCAM-1 positive) cells were sorted on BD Influx Cell Sorter (BD Biosciences) with 100 µm nozzle to PBS + 0.04% BSA. Single-cell cDNA libraries were prepared using the 10X Genomics Chromium Single Cell 3' reagent v2 chemistry in accordance with manufacturer's recommendations.