GSM2790902: Subcortex_P10; Mus musculus; RNA-Seq
After removal from the mother, embryos were stored on ice in Leibovitz`s L-15 Media and 1% Fetal Bovine Serum. Brains were removed from the embryos and imbedded in 1% UltraPure Low Melting Point Agarose and sectioned in 50-micron sections with a vibratome (Leica VT1200S). The CGE, MGE, cortical or subcortical region was dissected from each embryo. Tissue from multiple embryos was pooled together before dissociation. Embryonic brain tissue pooled from multiple embryos was dissociated into a single cell suspension using Papain dissociation system (Worthington Biochemical Corporation) according to the manufacturer’s instructions. Postnatal brain tissue was dissociated with 1mg/mL of pronase (Roche, #10 165 921 001) in ice cold prebubbled aCSF (Artificial Cerebrospinal Fluid) for 25 minutes. Fluorescent cells from Lhx6(BAC)-GFP brain tissue were sorted on a Sony SY3200 sorter with a 100 micron nozzle. Cells were sorted in bulk. The Chromium Single Cell 3' Library & Gel Bead Kit v2 (PN- 120237), Chromium Single Cell 3' Chip kit v2 (PN-120236) and Chromium i7 Multiplex Kit (PN-120262) were used according to the manufacturers instructions in the Chromium Single Cell 3' Reagents Kits V2 User Guide.