GSM7720311: C57WT48hPostONCRep2; Mus musculus; RNA-Seq
The eyeballs were enucleated and kept in ice-cold PBS. The retinas were quickly and carefully dissected, and whole retinal cells were dissociated using the Papain enzyme system (LK003150, Worthington). 30mM actinomycin-D was added to the Papain system to prevent post-enucleation transcription. The tissues then underwent manual trituration in neurobasal medium supplemented with 3% BSA, 10 mg/mL Ovomucoid, 1U/uL RNase inhibitor, and 3mM actinomycin D. Cells were filtered through a 40-μm strainer and centrifuged under 500´g. For scRNA-seq, the cell concentration was kept at ~1000 cells/uL. The scRNA-seq libraries were prepared using the 10X Genomics Chromium 3' Single-Cell Gene Expression V3 Kit following the standard protocol by the manufacturer. Approximately 8,000-12,000 cells were loaded onto the platform. Libraries were sequenced with the NovaSeq S2 100 platform.