title: GSM7683128: 4 weeks_maternal control diet_ZT21_replicate 1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from liver tissues using the E.Z.N.A. Total RNA Kit II (OMEGA, R6934). Briefly, 500 µL of RNA-Solv Reagent was added to approximately 50 mg of frozen liver tissue. We then added 3-4 mm zirconia beads. This mixture was subjected to homogenization in two 30-second cycles, interspersed with a 10-second interval, at 4°C using a KZ-III-F homogenizer (Servicebio). Following this, 200 µL of chloroform was added, and the solution was vigorously shaken for 20 seconds. The mixture was then allowed to settle for 3 minutes at room temperature before being centrifuged at 12,000 x g for 15 minutes at 4°C. The resulting RNA supernatant was collected and subsequently purified using the columns provided with the kit, in accordance with the manufacturer's protocol. RNA integrity of all samples was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies). Only RNA with a RIN above 8.0 were further processed. mRNA was purified from total RNA (3 μg of total RNA) using poly-T oligo-attached magnetic beads. Sequencing libraries were obtained using the NEBNext UltraTM RNA Library Prep Kit for Illumina following the manufacturer's instructions.