GSM1963126: FACS sorted brain mural cells 3; Mus musculus; RNA-Seq
We isolated double positive cells from the brain of Pdgfrb-eGFP/NG2-DsRed transgenic mice using FACS. Cells were lysed, RNA extracted and sequenced with next-generation sequencing (NGS). For comparison, we also determined the transcriptome of brain microvascular fragments (containing both endothelial cells and mural cells) isolated by mechanical tissue disintegration, collagenase digestion and immune-panning using anti-CD31 antibodies coupled to magnetic beads. RNA integrity and concentration of the FACS sorted cells and the isolated microvasculature was checked with the Agilent RNA 6000 pico kit and an Agilent 2100 Bioanalyzer (Agilent Biotechnologies). For conversion of the RNA to a cDNA library for Illumina sequencing, the SMARTer® Stranded Total RNA Sample Prep Kit - Low Input Mammalian kit from Clontech was used according to the manufacturer’s protocol. The total RNA was depleted from rRNA and tRNA’s with the RiboGone- Mammalian kit (Clontech) prior to library construction.
GEO Accession
GSM1963126