GSM7916528
Retinas of the vGlut2-Cre;LSL-Sun1GFP mice were quickly and carefully dissected out from enucleated eyeballs. Through gentle pipette trituration, the retinas were dissociated in ice-cold neurobasal medium on ice. According to the 10X Genomics protocol, whole retinal cells were then incubated in the nuclear lysis buffer on ice for 3-min to expose and penetrate the cell nuclei, and then centrifuged to pellet the nuclei. The cell nuclei were then resuspended in ice-cold MACS buffer (Miltenyi Biotec, Gaithersburg, MD) for anti-GFP MACS microbeads incubation. The whole retina nuclei were then passed through and eluted from the Miltenyi MACS MS column to enrich GFP+ RGC nuclei, then being centrifuged to pellet. The pellet was then resuspended to reach concentration of 4000~5000 nuclei/uL loading onto the 10X Genomics Single-Nuclei 3' HT platform. 10X Genomics Single-Nuclei 3' HT