GSM4314535: brainstem_10x; Homo sapiens; RNA-Seq
To dissociate brainstem organoids into single cells, they were incubated for ~30min in Accutase (STEMCELL TECHNOLOGIES) at 37°C. Droplet-based scRNA-seq libraries were generated using the ChromiumTM Single Cell 3' Reagent kits V2 (10x Genomics). Briefly, cell number and cell viability were assessed using the Countess II Automated Cell Counter (ThermoFisher). Thereafter, cells were mixed with the Single Cell Master Mix and loaded together with Single Cell 3' Gel beads and Partitioning Oil into a Single Cell 3' Chip. RNA transcripts were uniquely barcoded and reverse-transcribed in droplets. cDNAs were pooled and amplified according to the manufacturer's protocol. Libraries were quantified by High Sensitivity DNA Reagents (Agilent Technologies) and the KAPA Library Quantification kit (KAPA BIOSYSTEMS). Libraries then were sequenced by Illumina Hiseq 2500 in rapid mode.
GEO Accession
GSM4314535