GSM9594023: Cortex, fresh control, sample 2; Mus musculus; RNA-Seq
Following group-appropriate treatment, each sample was homogenized in TRIzol (Thermo Fisher Scientific) according to the manufacturer's instructions using a hand-held tissue homogenizer (Qiagen). Samples were subsequently shipped to the Beijing Genomics Institute (BGI) on dry ice for RNA extraction and RNA-seq. RNA libraries were prepared using the DNBSEQ Eukaryotic Stranded Transcriptome library preparation workflow. mRNA was enriched using oligo dT beads to capture poly(A)+ RNA. The enriched RNA was fragmented, and first-strand cDNA was synthesized using random N6 primers, followed by second-strand cDNA synthesis with dUTP replacing dTTP. The cDNA then underwent end repair, 3′ adenylation, and adaptor ligation. Before PCR amplification, the dUTP-marked strand was selectively degraded by Uracil-DNA-Glycosylase (UDG), and the remaining strand was amplified to generate a strand-specific cDNA library. The PCR products were heat denatured, circularized by splint oligo and DNA ligase, converted to DNA nanoballs, and sequenced on the DNBSEQ platform