GSM9040073: XY at D6 under testicular condition; Mus; RNA-Seq
Differentiated cells cultured under each condition were dissociated by incubation with 0.05% trypsin/EDTA for 7 minutes at 37 °C, followed by gentle pipetting with an equal volume of DMEM supplemented with 10% FBS, 2 mM glutamine, and 1× penicillin-streptomycin. The resulting single-cell suspension was incubated with BV421-conjugated anti-hCD271 antibody, and hCD271-positive cells were subsequently sorted using a BD FACSAria Fusion. The sorted cells were centrifuged and resuspended in XXXXX buffer. cDNA libraries were constructed using the Chromium Single Cell 3ʹ v3 Reagent Kits (10x Genomics). FACS-sorted cells were mixed with the Single Cell Master Mix to generate gel bead-in-emulsions (GEMs). Within each GEM, RNA was reverse-transcribed, and the resulting cDNAs were amplified by PCR. Sequencing libraries, adapted for the Illumina platform, were then prepared from the amplified cDNAs. The single-cell RNA-seq libraries were sequenced using paired-end reads on an Illumina HiSeq X Ten platform.