GSM9064486: 35 days post-booster replicate 2; Mus musculus; RNA-Seq
A total of six male mice were used in scRNA-seq experiments, with prostates pooled from three mice for each scRNA-seq sample. All four lobes of prostates were extracted, pooled, and then digested in complete RPMI with 1mg/ml Collagenase I on a shaker at 225 RPM and 37°C for 1.5 hours. Samples were filtered through a 70μm strainer, centrifuged at 250g for 5 minutes and resuspended. Dead cells were removed using the EasySep Dead Cell (Annexin V) Removal kit (STEMCELL Technologies, Vancouver, Canada) per the manufacturer's protocol. Cells were counted with trypan blue and diluted to a concentration of 1000 cells/μl, then prepped and loaded into the 10x Chromium (10x Genomics, Pleasanton, CA) instrument. Manufacturer's recommended protocol for a recovery of upto 10000 cells were followed.