GSM4720584: e12.5 cervical motor neurons; Mus musculus; RNA-Seq
Spinal cords from cervical segment were dissected in L15 medium and cells were dissociated using enzymatic (papain) and mechanical approach, following manufacturer's instruction (Neural Tissue Dissociation Kit (P) and gentleMACS dissociator, Miltenyi Biotec). Cell suspension was filtered through 70µm strainer and subjected to FACS under 45 psi to enrich GFP-positive cells . Collected cells were centrifuged at 1000rpm for 5 minutes and resuspended in 1%FBS supplemented DMEM. Cells viability and concentration was determined using Countess II Automated Cell Counter. Cells were loaded into 10x Chromium Controller using Single Cell 3' v3.1 reagents for single cell isolation. scRNA-seq libraries were generated following manufacturer's instruction (10X Genomics, Chromium Single Cell 3' v3.1) and sequenced using a NextSeq 500/550 platform with pair-ended reads (PE150: Read 1: 28 bp; Read 2: 122 bp). A total of 6193 cells were sequenced up to 79k mean reads per cell.
GEO
GSM4720584