GSM3418163: fWBI Animal 5, White Matter; Macaca mulatta; RNA-Seq
Animals were humanely euthanized in accordance with the American Veterinary Medical Association's Guidelines on Euthanasia by deep anesthesia with pentobarbital, followed by exsanguination and perfusion of the vascular system with 2 liters of cold normal saline. The brain was removed intact, and sectioned coronally in 4 mm intervals using a stainless steel brain matrix with cutting guides. Sections were immediately frozen on dry ice. 100 mg of brain tissue was homogenized in QIAzol lysis reagent, and aliquots of homogenized lysates equivalent to 40 mg tissue were extracted for total RNA using the RNeasy Microarray Tissue Mini kit (Qiagen). Extracted RNA was DNase-treated and purified using the RNA Clean and Concentrator-5 kit (Zymo Research), then assessed for RNA quality using an Agilent 2100 Bioanalyzer and the RNA 6000 Nano Kit (Agilent Technologies). RIN values for the RNA samples ranged from 7.1 to 9.3. Total RNA was used to prepare cDNA libraries using the Illumina® TruSeq Stranded Total RNA with Ribo-Zero Gold Preparation kit (Illumina Inc.) and the SciClone NGS Work Station (Perkin Elmer).