GSM6407435: Tsc1 RPE KO, scRNAseq; Mus musculus; RNA-Seq
After removal of the neuroretina, the posterior eye cups were digested with the mixture of collagenase and dispase in the presence of LIVE/DEAD fixable near-IR dead cell dye for 15 min at 37°C.  After washing, the cells were digested with 0.05% Trypsin for 5 min at 37°C.  The cell suspension was pelleted, and washed with resuspension buffer (PBS containing 0.04% BSA).  After removing the cell clumps with a 40 m cell strainer, the single-cell suspension was sorted via FACS with gates set to include live cells.  Collected cells were used to prepare barcoded, next-generation sequencing libraries of single cells. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics).