GSM7916099: MECP2T203M; Homo sapiens; RNA-Seq
Briefly, 5-7 cerebral organoids were dissociated into a single-cell suspension via incubation with 1 mL tryp-LE (Life Technologies, Thermo Fisher Scientific) for 35 minutes at 37°C with gentle agitation every 5–8 minutes, followed by 3 washes with 2% FBS in DPBS and gentle titration using a P200 pipette. Cell suspension approximately 10,000 cells was loaded onto a Chromium Single Cell 3' controller (10x Genomics) to generate single-cell gel beads in the emulsion (GEMs) according to the manufacturer's protocol. According to the manufacture's introduction, Single-cell RNA-seq libraries were constructed using Single Cell 3' Library and Gel Bead Kit V3.1 (10x Genomics). The libraries were then sequenced using an Illumina Novaseq6000 sequencer with a sequencing depth of at least 100,000 reads per cell with pair-end 150 bp (PE150) strategy.