title: GSM5969879: ONCd7C57RGCImmuneAstro2_S14_L007; Mus musculus; RNA-Seq
library_construction_protocol: Retinas were dissected in oxygenated AMES, digested in papain, and dissociated to single cell suspensions using manual trituration in ovomucoid solution. Eyecups were dissected in HBSS+HEPES and digested in trypsin/EDTA, after which RPE sheets were released by manual shaking into 20% FBS in HBSS+HEPES, and dissociated into single cell suspensions using manual trituration. Single cell suspensions were incubated with anti-CD90, CD45, CD140a and GLAST antibodies (conjugated to various fluorophores) for 10 million cells per 100µl. Cells were sorted using a MoFlo Astrios. Cellular debris, doublets, and dead cells were excluded. Collected cells were loaded into 10X Chromium Single Cell Chips at ~1,000 cells/ul. Single cell libraries were made with Chromium 3' V2 or V3 platforms (10X Genomics, Pleasanton, CA) following the manufacturer's protocols. On average, approximately 10,000 single cells were loaded on each channel and approximately 6,000 cells were recovered. Libraries were sequenced on Illumina NextSeq 500 or HiSeq X.