GSM4039247: M-tumor-1; Mus musculus; RNA-Seq
Animals were perfused with PBS and brains taken out. Brain tissue was enzymatically dissociated to single cell suspension with Neural Tissue Dissociation Kit [Miltenyi Biotec] and gentleMACS Octo Dissociator [Miltenyi Biotec]. Myelin was removed by density gradient centrifugation and CD11b+ cells FACS sorted to 20% FBS in PBS. Cell suspension from FACS was loaded directly onto Chromium™ Single Cell A Chip [10x Genomics]. Preparation of Gel Beads in Emulsion and libraries were performed with Chromium Controller and Single Cell Gene Expression v2 Chemistry [10x Genomics, USA] according to the Chromium Single Cell 3' Reagent Kits v2 User Guide provided by the manufacturer. Libraries quality and quantity was verified with High Sensitivity DNA Kit [Agilent Technologies, USA] on 2100 Bioanalyzer [Agilent Technologies, USA]. Next, sequencing was run in the rapid run flow cell and paired-end sequenced (read 1 – 26 bp, read 2 – 100 bp) on HiSeq 1500 [Illumina, USA]