title: Single cell RNA sequencing of 120h after aggregation mouse gastruloids cultured with different doses of retinol
design: Single cell RNA sequencing of 120h after aggregation mouse gastruloids cultured with different doses of retinol
library_construction_protocol: Gastruloids were collected and washed with ice-cold PBS three times. They were incubated with TrypLE Express (Gibco, 12605028) for 2X 4 min in a heating block at 37oC, shaking at 300 rpm. After incubations, the gastruloids were dissociated into single cells by pipetting up and down 10 times. The cells were washed with PBS twice, centrifuged at 1300 rpm for 5 min at 4ì§¸C, and the supernatant was carefully discarded without disturbing the pellet. The cells were resuspended in PBS containing 0,04 % BSA (Sigma-Aldrich, A3059) and 1:5000 DAPI (Invitrogen, D1306) as a viability dye. The cell suspension was filtered through a FACS  (Falcon, 352235), and FACS analysis was performed (CytoFlex SRT Beckman Coulter). DAPI-negative and singlet populations were gated and collected. 10X Chromium Genomics workflow was performed by polyA enrichment (3'). 10X Chromium Genomics workflow was performed by polyA enrichment with single cell resolution.
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-15522:Sample 1_p