GSM8831236: P6 Ki67CreERT/+;Rosa26RtdTomato/+ cochlea, rep2; Mus musculus; RNA-Seq
Following microdissection, cochlear tissues were incubated in 0.5mg/mL Thermolysin at 37°C for 45 minutes, followed by incubation in Accutase for 20 minutes. The reaction was stopped by adding 300 µL of DMEM/F12 with 5% fetal bovine serum, and the tissue was titurated 60x by pipetting to dissociate into single cells. Cell clumps were removed by filtering the cells through a 40 µm pluriSelect filter mini strainer, followed by pelleting in a 4C centrifuge for 5 min at 300 rcf. Supernatant was aspirated and the cell pellet was resuspended in 60 µL DMEM/F12 with 5% FBS. After quantifying cell numbers and viability, cells were placed on ice and brought to the Stanford Functional Genomics Core for 10x Genomics scRNA-seq capture and sequencing. 10x Genomics Single Cell 3' v3