scRNA-Seq of mus musculus: adult female spinal cord
A spinal cord, dissected from each female mouse transcardially perfused with PBS, was dissociated in Neural Tissue Dissociation Kit-Postnatal Neurons reagents by using gentleMACS Dissociator (Miltenyi Biotec, Germany) at 37 C for 15 min. To remove myelin debris, the cells were incubated with Myelin Removal Beads II (Miltenyi Biotec, Germany) according to the manufacturers instruction and then purified by autoMACS Pro Separator (Miltenyi Biotec, Germany). The living single-cells were sorted by FACSMelody Cell Sorter (BD Biosciences, USA) after 7-AAD (7-Aminoactinomycin D) staining. The barcoded libraries were generated by Chromium controller (10x genomics, USA) using Chromium Next GEM Single Cell 3 Reagent Kits v3.1 for dual index and sequenced with 150-nt paired-end reads by using Illumina Hiseq 2500 (Illumina, USA).