title: GSM8009845: sequenced matrices from brain_KO_day50; Mus musculus; RNA-Seq
library_construction_protocol: For dissociation of brain and spinal cord tissue for single cell sequencing, 2 ml glasspipette tips was used tohomogenize the tissue and dissociated with papain (Worthington) supplemented with DNAse in Ca2+-free serum-free Earle's Balanced Salt Solution pre-heated to 37C.Tissue dissociated at 37C for 25 min, with homogenization after 10 minutes using 1 mL plastic tip. After 25 min the tissue was homogenized again with 1 mL plastic tip andpassed through 70 um filter and Dulbecco's Modified Eagle Medium (Gibco) supplemented with 10% Fetal Bovine Serum added, spun at 300g for 10 mins. Cells subjected to myelin removal beads (Miltenyi), counted, diluted in Ca2+ -free Hank's Balanced Salt Solution with 0.5% Bovine Serum Albuminand loaded into a Chromium Single Cell 3' Chip (10X Genomics) and processed follow manufacturer's instructions. Sequencing libraries were prepared with the Chromium Single Cell 3' library and Gel Bead kit v2 (10X Genomics). Libraries were sequenced on a Nova-seq 6000.Reads were aligned to the mouse genome assembly GRCm38.