GSM9788201: HSC_TGFB_Res_rep3; Homo sapiens; RNA-Seq
Total RNA was extracted from liver tissue using the TRIzol–chloroform method following the manufacturer's protocol. RNA quality and quantity were assessed using a NanoDrop spectrophotometer (selecting samples with A260/A280 ratios between 1.8 and 2.0). RNA integrity was subsequently confirmed (RIN ≥ 8) prior to library preparation. DNBSEQ Eukaryotic Strand-specific mRNA library. Total RNA samples were enriched for polyA mRNA using oligo dT beads, followed by mRNA fragmentation. Fragmented mRNA was synthesized into first-strand cDNA using random primers. Second-strand cDNA synthesis was performed incorporating dUTP instead of dTTP to preserve strand specificity. The synthesized cDNA was subjected to end-repair, 3' adenylation, and adaptor ligation before PCR amplification. After library quality control and circularization, the library was amplified to generate DNA nanoballs (DNB) and sequenced on the DNBSEQ platform with a PE100 (paired-end 100bp) strategy.