title: GSM8969044: C57BL/6J, Olfactory epithelium, total RNA, OC condition, OC Mixture 2; Mus musculus; RNA-Seq
library_construction_protocol: Olfactory epithelium tissues were rapidly collected from mice immediately after odorant exposure and perfused with chilled buffer (PBS, 5mM NaF, 2.5mM Na3VO4, 2.5mM Na4P2O7, 5mM β-glycerophosphate, 100μg/mL cycloheximide, protease inhibitor). The tissues were homogenized in a buffer containing 10mM HEPES (pH 7.4), 150mM KCl, 10mM MgCl2, 100mM calyculin A, 0.5mM DTT, 100U/mL RNasin, 250μg/mL cycloheximide, and protease inhibitors using a TissueLyser II (Qiagen) for 2 minutes at 20Hz. Homogenates were centrifuged at 2,000×g for 10 minutes at 4°C. The supernatant was supplemented with 0.7% NP-40, incubated on ice, and centrifuged at 20,000×g for 10 minutes at 4°C. A 50μL aliquot of the olfactory epithelium homogenate was mixed with 350μL of Buffer RLT (Qiagen) and incubated on ice for 5 minutes. Total RNA was purified using the RNeasy Mini Kit (Qiagen) according to the manufacturer's protocol, including on-column DNase digestion. Libraries were prepared using the TruSeq Stranded mRNA Sample Prep kit (Illumina), and sequencing was carried out on an Illumina NovaSeq 6000 platform according to the manufacturer's protocols.