GSM9372059: Liver RNA-seq HSC-PCK1-KO 3 (87); Mus musculus; 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. Messenger RNA (mRNA) was enriched from total RNA using oligo(dT) beads. The enriched mRNA was fragmented into small pieces and reverse-transcribed into first-strand cDNA using random primers. Second-strand cDNA synthesis was performed using dUTP to achieve strand specificity. The cDNA fragments underwent end-repair, 3'-adenylation, and adaptor ligation. After PCR amplification and quality control , the library products were circularized to generate DNA Nanoballs (DNBs). Sequencing was performed on the DNBSEQ platform with a PE100 (paired-end 100bp) strategy.