GSM6755407: C_wt; Mus musculus; RNA-Seq
Total RNA of one cortical hemisphere was extracted using TRIzol™ Reagent (Invitrogen™) and chloroform (Sigma Aldrich), followed by centrifugation at 12000 g for 15 min at 4°C. The upper phase was transferred to a fresh tube and 1.5 volumes of 100% EtOH were added. Total RNA was purified by using the RNA Clean&Concentrator-5 prep Kit (Zymo Research). The samples were further treated with RQ1 RNase-Free DNase (Promega) as described in the kit instructions manual. RNA concentration and quality was assessed by using the NanoDrop spectrophotometer (Thermo Fisher Scientific) and the Bioanalyzer 2100 with the RNA 6000 Nano kit (Agilent). cDNA libraries were generated with the SENSE mRNA-Seq Library Prep Kit V2 (Lexogen) using 1.5 µg total RNA. The quality of the generated libraries was monitored by using the High Sensitivity DNA Analysis Kit (Agilent) and the Bioanalyzer 2100. Libraries were sequenced on an Illumina HiSeq 2500 instrument.