GSM4745562: Rat 4_hypothalamus_Ozone_Female; Rattus norvegicus; RNA-Seq
Fetal and placental endpoints were collected for assessment at GD 21. Dams and fetuses were euthanized using a lethal intraperitoneal dose of pentobarbital (> 400 mg/kg). For a subset of fetuses in each litter (1st and 2nd fetus on the ovarian side of each horn), the hypothalamus was dissected, quick frozen in liquid nitrogen,and stored at -80ºC until use. Total RNA was isolated from fetal whole hypothalamus using Trizol with the Direct-Zol RNA Purification Kit (Zymo Research). Isolated RNAs were quantified by the Qubit fluorometer (ThermoFisher) and quality checked by Bioanalyzer (Beckman Coulter, Inc.; Santa Clara, CA). All samples had a RIN at or above 9. Hypothalamic RNA samples (n = 6/sex/group from the same subset of selected fetuses, 1 μg each) were processed on Apollo324 system (Takara Bio USA, Inc.; Mountain View, CA) using PrepXTM mRNA selection and mRNA-seq library prep for Illumina. The products from library preparation were amplified by PCR with the sequencing index primers. The PCR products were purified on Apollo324 and assessed for quality using Bioanalyzer. The library concentrations were verified by Qubit.