GSM9407190: Control HH3/3plus stage embryo with 5h additional incubation; Gallus gallus; RNA-Seq
Embryos were dissected in cold PBS+/+, and approximately half of the area opaca was trimmed to enrich for embryonic tissue. Each embryo was transferred to a 1.5mL Eppendorf tube, washed 3× with PBS+/+, and incubated in 1 mL 0.05% trypsin (Thermo Fisher Scientific) at 37C for 15min with gentle trituration every 5min. Trypsin was quenched with 100uL FBS, and cells were pelleted (450×g, 5min, 4C) and resuspended in PBS + 0.05% BSA (Sigma-Aldrich). Cell viability (85–95%) was assessed via haemocytometer. Single-cell suspensions were submitted to the CRUK CI Genomics Core Facility for 10X Genomics library preparation and Illumina sequencing.