GSM9700031: PA_3-IAA_rep2; Bos taurus; RNA-Seq
Total RNA was extracted from bovine hepatocytes using RNAiso Plus (D9108, TaKaRa) according to the manufacturer's instructions. RNA concentration and purity were determined using a NanoDrop 2000 spectrophotometer, and integrity was assessed by agarose gel electrophoresis. RNA quality was further evaluated using an Agilent 5300 system to obtain the RQN value. Only samples with total RNA ≥ 1 μg, concentration ≥ 30 ng/μL, RQN ≥ 6.5, and an OD260/280 ratio between 1.8 and 2.2 were used for library construction. mRNA was enriched from total RNA using oligo(dT) magnetic beads and fragmented into ~300 bp fragments. Double-stranded cDNA was generated through reverse transcription and second-strand synthesis. The cDNA fragments were end-repaired, A-tailed, and ligated to adapters, followed by size selection and PCR amplification to construct sequencing libraries. The libraries were sequenced on  Illumina Novaseq X Plus following the vendor's recommended protocol.