title: GSM9085913: DRGs, Penk ctrl, pooled; Mus musculus; RNA-Seq
library_construction_protocol: Cervical region of spinal cord was isolated from mice and immediately fixed in 4% PFA overnight at 4 °C. Following PBS washes, dorsal root ganglia (DRGs) from spinal levels C2 to C7 were bilaterally dissected. Tissue from each mouse was pooled and mechanically homogenized in 1 ml of pre-warmed RPMI 1640 medium containing collagenase I (3000 U/ml; Worthington Biochemical Corporation) for 20 minutes at 250 rpm and 37 °C in a shaking incubator. Additional dissociation was performed using a Dounce homogenizer at room temperature until the majority of DRGs were dissociated. The resulting cell suspension was filtered through a 70 µm mini cell strainer (pluriSelect #43-10070-40) and centrifuged at 850 g for 5 minutes at room temperature. The supernatant was discarded, and pellet was resuspended in 1 ml of quenching buffer (10x Genomics, PN 2000516). Library preparation was performed using the Chromium Next GEM Single Cell Fixed RNA Profiling Kit for multiplexed samples, following the manufacturer's protocol (Mouse Transcriptome Kit #PN-1000496, Chromium Next GEM Chip Q Single Cell Kit #PN-1000418, and Dual Index Kit TS Set A #PN-1000251). Reads were aligned to the mouse reference genome (GRCm39) using Cell Ranger v7.0 (10x Genomics).