GSM9677522: PAH_sham_1; Rattus norvegicus; RNA-Seq
Lung tissues were collected for total RNA extraction using TRIZOL reagent (15596-018, Ambion, USA) according to the manufacturer's protocol. RNA quality and concentration were assessed using an Ultrafine spectrophotometer (N50 Touch, IMPLEN, Germany), and RNA integrity was confirmed by 1.0% agarose gel electrophoresisFor each sample, 1 μg of total RNA was treated with RQ1 DNase (M6101, Promega, USA) to eliminate residual genomic DNA. RNA sequencing was conducted by Wuhan Ruixing Biotec Co., Ltd. For each sample, 1 μg of total RNA was treated with RQ1 DNase (M6101, Promega, USA) to remove residual DNA. Strand-specific RNA-seq libraries were constructed using the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina (NR605, Vazyme, China). mRNA was enriched with VAHTS mRNA Capture Beads (N401, Vazyme, China), followed by fragmentation and synthesis of double-stranded cDNA. After end repair, A-tailing, and ligation to VAHTS RNA Multiplex Oligos Set 1 for Illumina (N323, Vazyme, China), the ligated products were amplified, purified, quantified and stored at –80°C until sequencing. The use of dUTP marking during second-strand synthesis enabled strand-specific sequencing. Libraries were sequenced on the Illumina NovaSeq Xplus platform following the manufacturer's instructions, generating 150 bp paired-end reads