title: GSM9103401: NMDA-24; Mus musculus; RNA-Seq
library_construction_protocol: Fluorescence activated cell sorting Retinae were dissected in ice-cold PBS and dissociated in 10 units Papain/60 units216 DNAse (500μL/30μL respectively for 3-6 retinae) for 25-30 min in a shaking water bath at 37oC. Dissociated retinae were triturated 10 times with a polished pipette and EBSS-Ovomucoid-DNAse was added to stop digestion. The cell suspension was filtered using a 40μm strainer and centrifuged at 900 RPM for 8-10 min. For single cell analysis, cell pellets were resuspended and kept in DNAse/Ovomucoid/Neurobasal (1:1:10 respectively) during cell sorting. Cell sorting was performed using the Aria III cell sorter (BD Biosciences). Tomato positive and negative cells were sorted using an 80-micron nozzle, captured in chilled tubes and resuspended in 0.04% ultrapure BSA in 0.1M PBS. Fresh cell suspensions were immediately subjected to the scRNA-seq protocol. FACS sorted TdTomato+ cells were counted and diluted in 1X PBS with 0.04 % BSA (Sigma Cat# SRE0036) before being loaded onto the 10X Genomics Chromium instrument. The scRNA-seq libraries were generated using the 10X Chromium Single Cell 3' v2 reagent kit according to the manufacturer's instructions and were sequenced on an Illumina Nextseq500.