GSM4878689: Control_PLX5622_2; Mus musculus; RNA-Seq
Mice  were  perfused  with  ice-cold  PBS,  brains were rapidly removed, and the cortex was microdissected. The cortex was dissociated using the Adult Mouse Brain Dissociation Kit for mouse and rat per manufacturer's instructions with minor modifications(Miltenyi  Biotec,Bergisch  Gladbach,  Germany). aCSF  (as  described  below)  was used  to  make  up  enzyme  solutions.  Dissected  cortex was  immediately  placed  into  pre-warmed (37°C) Enzyme P solution, Enzyme A solution was added, and tissue was dissociated in C-tubes at  37°C  using  a  gentleMACS  dissociator(Miltenyi  Biotec).  Density  centrifugation  was  used  to remove myelin debris, remaining red blood cells were lysed, and cell viability and concentration was determined using a hemocytometer. Approximately 3,000-7,000 cells were recovered from each cortical homogenate. Single-cell cortical suspension was loaded onto a 10X chip  and  run  on  a  Chromium  Controller  to  generate  gel-bead  emulsions  (10X  Genomics, 12Pleasanton, CA, USA). Protocol for UMI barcoding, cDNA amplification, and library construction followed the 10X Genomics Single Cell 3' v2 reagent kit protocol. Library quality was determined by Agilent High Sensitivity DNA BioAnalyzer chip. RNA-sequencing was performed using an Illumina HiSeq by Novogene per 10x Single Cell 3' v2 specifications