GSM6387314: 72h_FGF2; Mus musculus; RNA-Seq
Gastruloids wrere washed twice in PBS (5 min at 1000 RPM) and dissociated to single cells by incubation with TrypLE Express (Gibco) for 10 min at 37C, with pipetting at 5min. Cells were washed with PBS (5 min at 1000 RPM) and passed through 70µm cell strainers and re-suspended in 1% BSA in PBS. Single cell suspensions were assessed for cell number and viability using an EVE automated cell counter (NanoEnTech) with trypan blue staining.  Approximately 10,000 cells were processed for single cell mRNA-Seq using the 10x Chromium 3' mRNA-Seq kit version 3.1 according to the manufacturer's instructions.  Sequencing was carried out on the HiSeq 4000 (Illumina) with a read configuration conforming to the required sequencing parameters for single indexed 10x 3' mRNA-Seq libraries. 10X_3prime_mRNA