title: GSM7310351: mouse brain, sham, rep3; Mus musculus; RNA-Seq
library_construction_protocol: The total RNA samples were treated with Trizol (Life Technologies, USA) in accordance with the kit's instructions. The total RNA concentrations and quality were assessed by NanoDrop-1000 and Agilent 2200. Specifically, OD260/OD280 remained between 1.8 and 2.2, and 28S/18S > 1.5, RIN >8, while the concentrations of total RNA were to be over 400 ng/μl. EpicentreRibo-Zero rRNA Removal Kit (Illumina, USA) was used to eliminate all rRNAs from the total RNA. Purified RNA fragments were subjected to the first strand and second-strand cDNA synthesis. Afterwards, adapter-ligated and enriched RNA-Seq libraries were prepared using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (NEB, USA). DNA was amplified. The library's quality and concentration were validated by Agilent 2200 TapeStation (Agilent Technologies, USA). The obtained libraries were subjected to paired-end sequencing on an Illumina HiSeq2500 platform.