GSM9305566: Three pooled e17.5 eye cups from Myrf humdG/humdG strain; Mus musculus; RNA-Seq
Eyes were collected from e17.5 litters, the cornea, lens, and optic nerve were removed, and the eyecups were place on ice in 1 X HBSS (Invitrogen, 14175095) while samples were genotyped using a rapid genotyping protocol (Truett et al., 2000). Three Myrffl/fl and three Rx>cre Myrffl/fl samples were pooled for each time point.  Eyecups were dissociated into single cells in a Papain solution containing 5mM L Cysteine (Sigma, C7352), 1mM EDTA (Sigma, E4884), 0.6 mM 2-mercaptoethanol (Sigma, M6250), and 1mg/ml Papain (Roche, 10108014001) for 10 minutes at 37°C with trituration every 2 minutes.  The dissociation was stopped using Neurobasal Media (Invitrogen, 12348017) with 10% Fetal Bovine Serum (FBS) (Corning, MT35010CV), single cells were collected by centrifugation for 5 minutes at 300 RCF at 4°C.  The supernatant was aspirated, and the single cells were resuspended in cold Neurobasal Media with 3% FBS. Single cell libraries were prepared by the University of Michigan Advanced Genomics Core and sequencing was performed using the 10X Genomics Chromium platform using manufacturer's protocol for 10x Single Cell Expression 3' and sequenced on the NovaSeq (S4) 300 cycle (Illumina, San Diego, CA).