GSM3614615: PFHT1_ROW36_2_R2; Mus musculus; RNA-Seq
Coordinated pregnant mice of 12 to 30 weeks were ethically sacrificed to extract E15.5 embryos eyes. Thirty retinas were extracted in ice-cold L15, micro-dissected under a stereomicroscope and incubated in 200µl single cell dissociation solution consisting in a papain (1mg/ml) enriched HBSS at 37°C for 12 minutes with trituration every 2 minutes. At the end of the incubation time, cells were further dissociated via gentle up-and-down pipetting. Reaction was stopped with the addition of 400µl of ovalbumin enriched cold HBSS and the cell suspension was then passed on a 40µm cell strainer to remove cellular aggregates.  Cells were then centrifuged 5 min at 500 G at 4°C. After removal of the liquid, the pellet was suspended in 250µl of cold HBSS and the resulting solution was finally FAC-sorted on a MoFloAstrios device (Beckman) to reach a concentration of 410 cells /µl. The libraries of single-cells were prepared using the SmartSeq2 platform following the manufacturer's protocol.