GSM8069908: S3_D5_18h, replicate 1, scRNAseq; Homo sapiens; RNA-Seq
16 (day7) to 32 (day3) notoroids were dissociated by first washing with PBS-/- and then incubating with accutase for 10min at 37°C following mechanical dissociation and another incubation at 37°C. Accutase suspension was then diluted in 4x volume with resuspension buffer (DMEM/F12 with 1% BSA). The cell suspension was then spined for 4min at 0.6xg, resuspended in 250µL of resuspension buffer and filtered through a 40µm Flowmi cell strainer (cat. no. 136800040) and twice through a pre-wet 20µm pluriSelect strainer (43-10020-60). The single-cell suspension was diluted to 1300 cells/uL and had a viability >95%. The samples were separately loaded for capture with the Chromium System using the Single Cell 3′ v3.1 reagents (10X Genomics). cDNA amplification involved 12 PCR cycles. Libraries for the samples were multiplexed so that the number of reads matched one lane per sample and sequenced on an Illumina HiSeq4000 using 100 bp paired-end runs.