GSM7416841: P21_Worthington_rep2; Mus musculus; RNA-Seq
Tissue pieces were processed to obtain a single cell suspension using papain digestion  (30-90   min   according   to   the   mice   age),   following   an   adapted   protocol   from   the  Worthington Papain Dissociation System kit  (Worthington Biochemical).  Single cells  were   resuspended   in   ice-cold   Hibernate-A   containing   10%   fetal   bovine   serum.  Oligodendrocytes were isolated by fluorescence activated cells sorting. Sorted cells  were spun down and resuspended into 0.04% BSA in PBS. Cell were counted in a  hemocytometer chamber and immediately processed for single cell GE formation. Library was performed according to the manufacter's instructions with the single cell 3'  v2 protocol (or v3 for the second replicate) (10x Genomics).  Approximately 6-8,000  sorted oligodendrocytes were loaded onto  a Chromium™ Single Cell 3' Chip and  processed   through   the   Chromium   Controller   to   generate   single-cell   gel   beads   in  emulsion.   cDNA   amplification   and   library   construction   were   done   following  10xGenomics instructions. Libraries were quantified in BioAnalyzer and sequenced  with a NextSeq 500 instrument (Illumina).