title: GSM4816782: Day10 EPS-blastoids; Homo sapiens; RNA-Seq
library_construction_protocol: EPS-blastoids were picked up by mouth pipette and washed with PBS containing 0.05% BSA. About 200 blastoids on day 6, 40 blastoids IVC structures on day 8 and 20 blastoids IVC structures on day 10 were collected for single-cell RNA-seq. Samples on day 6 were dissociated with enzyme mix composed of 0.5×Versene (Lonza, 17711E), 0.5×Accumax (STEMCELL Technologies, 07921) and 0.05×DNaseI (STEMCELL Technologies, 07900) at 37°C for 30 min with agitation and terminated by 5% BSA in PBS. Samples on day 8 or 10 were dissociated with enzyme mix composed of 0.25% Trypsin (Thermo Fisher Scientific, 25200056), and 0.05×DNaseI (STEMCELL Technologies, 07900) at 37°C for 15 min with agitation and terminated by 5% BSA in PBS. Dissociated cells were repeated pipetting and washed with PBS containing 0.05% BSA. Dissociated cells were repeated pipetting and washed with PBS containing 0.05% BSA. Single-cell RNA-seq libraries were constructed using Single Cell 3' Library and Gel Bead Kit V3(10×Genomics, 1000075). The cell suspension (300-600 living cells per microliter determined by Count Star, about 20,000 dissociated cells each sample) were loaded onto the Chromium Single Cell B Chip (10×Genomics, 1000074) and processed in the Chromium single cell controller (10×Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol.  Finally, sequencing was performed on the Illumina Novaseq6000 sequencer with a sequencing depth of at least 60,000 reads per cell and 150 bp (PE150) paired-end reads (performed by CapitalBio, Beijing).