title: GSM8371506: dorsal root ganglia, AEW, rep2; Mus musculus; RNA-Seq
library_construction_protocol: L4-5 DRGs from adult mice (8 weeks) were collected in cold complete saline solution (CSS, 137 mM NaCl, 5.3 mM KCl, MgCl2-6H2O, 25 mM Sorbitol, 10 mM HEPES, 3 mM CaCl2, pH=7.2) and then incubated in an enzyme solution in CSS composed of 0.35 U/ml Liberase TM (Roche) and 0.6 mM EDTA TM 37 °C for 20 minutes, followed by incubation at 37°C for 10min in CSS composed of 0.35 U/ml Liberase TL (Roche), 0.6 mM EDTA, and 30 U/ml papain (Worthington Biochemical). Enzymatic digestion was stopped by adding 1ml of DRG media supplemented with 1.5 mg/ml bovin serum albumin (BSA, Sigma) and 1.5 mg/ml trypsin inhibitor (Sigma). After gentle trituration with 1 ml pipette, DRG neurons were centrifuged at 800rpm for 2-3 min and resuspended in 1 ml DH10 solution (90% DMEM/F-12, 10% FBS, 100 U/ml penicillin, and 100 μg/ml Streptomycin, Gibco). RNA libraries were generated by 10xGenomics Single Cell 3' V3.1 Library Gel Bead Kit and prepared for sequencing using standard Illumina protocols