title: GSM8484756: Mouse, OPC-ITAN & OPC-LacZ, 20-28W, GEX, multiplexing; Mus musculus; RNA-Seq
library_construction_protocol: Four mice IDH-mutant glioma models (OPC-ITAN, 26 to 38-weeks-old-age) and a control mouse (OPC-LacZ, 20-weeks-old-age) were euthanized by trans-cardiac perfusion with 20ml cold DPBS without Ca2+/Mg2+ under isoflurane anesthesia. GFP-positive areas in brain were micro-dissected with a scalpel blade and pooled for digestion using the Adult Brain Dissociation Kit, mouse and rat (#130-107-677, Miltenyi Biotec), kept cold on ice. After manual gentle tissue dissociation, cell debris in the single cell suspension were removed by a gradient centrifugation (3000×g for 10min) at 4℃. The single cell suspension was labeled with immune cell markers, for example, CD11b-APC (1:100 dilution; #101212, BioLegend) and CD45-APC-Cy7 (1:100 dilution; #103116, BioLegend), in flow cytometry buffer. DAPI (0.5μg-1; #422801, BioLegend) stained single cell suspension, to discriminate dead cells from live cells, were isolated with FACSAria Fusion (BD). Except for the "mouse4", all samples were labled with CellPlex kit (10X Genomics). RNA libraries for single-cell RNA-seq Chromium Single Cell 3' kit v3.1 (10X Genomics) following manufacturer's instructions.