title: GSM7777504: Tcell_depletion_biological_replicate_1; Mus musculus; RNA-Seq
library_construction_protocol: For 10X genomic experiments, mice were deeply anesthetized and perfused with cold PBS. Each brain was removed and individually microdissected under a dissection microscope, wherein the cortex and hippocampus were isolated from the brain and the attached choroid plexus was removed. Cells were isolated with a previously established isolation protocol 27using gentleMACS with the Neural Tissue Dissociation Kit (Papain; Miltenyi Biotec) and actinomycin D (Act-D, Sigma-Aldrich, catalog no. A1410) at a final concentration of 45 mM. After dissociation, myelin debris were removed from the cell suspension using Myelin Removal Beads II (Miltenyi Biotec). The cells were resuspended in 0.04% BSA+PBS and cells were counted using automated cell counter (TC20 Bio-Rad) before loading to Chromium Controller. Chromium Single Cell 3′ Library & Gel Bead Kit v.3.1 (10X Genomics) Single-cell suspensions were loaded on to the Chromium Single Cell Controller using the Chromium Single Cell 3′ Library & Gel Bead Kit v.3.1 (10X Genomics) chemistry following the manufacturer's instructions. Sample processing and library preparation were performed according to the manufacturer's instructions using AMPure beads (Beckman-Coulter). Libraries were sequenced on the DNBSEQ Sequencing System (Dresden).