GSM9347057: Cochlea, CBA/CaJ, 16W, mock 3; Mus musculus; RNA-Seq
For Bulk RNA-sequencing,total RNA was extracted from cochlear tissue using TRIzol reagent (Thermo Fisher) according to the manufacturer's protocol, and further purified with the RNeasy Plus Mini Kit (Qiagen). Purified RNA was sent to Novogene (Sacramento, CA, USA) for quality control (QC), library preparation, sequencing, and quantitation according to their standard gene expression workflow. RNA degradation and contamination were initially assessed on 1% agarose gels. RNA purity was evaluated using a NanoPhotometer spectrophotometer (IMPLEN, CA, USA), while RNA integrity and concentration were determined using the RNA Nano 6000 Assay Kit with the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). Only RNA samples that passed QC were used for library preparation and sequencing. For library construction, 1 µg of total RNA per sample was used as input. Libraries were prepared with the NEBNext Ultra™ RNA Library Prep Kit for Illumina (NEB, USA) according to the manufacturer's instructions, with unique index codes added to each sample.