GSM9453138: Saline_PBMC_biol_rep5; Mus musculus; RNA-Seq
Peripheral blood mononuclear cells were isolated by density-gradient centrifugation. Total RNA was extracted from mouse PBMCs using the Illumina TruSeq RNA Sample Prep V2 Kit (#RS-122-2001). Library preparation and RNA sequencing were conducted by Beijing Novogene Corporation. Messenger RNA underwent poly(A) selection from ~1 μg total RNA via oligo(dT)-coupled magnetic beads, followed by fragmentation and first-strand cDNA synthesis. Double-stranded cDNA was generated, ligated with barcoded adapters using the VAHTS Universal V6 RNA-seq Library Prep Kit (#NR604-02, Vazyme), and quantified via Qubit 2.0 Fluorometer (Thermo Fisher). Library fragment size distribution was assessed using an Agilent 2100 Bioanalyzer. Paired-end sequencing (2 × 150 bp) was performed on the Illumina NovaSeq 6000 platform, yielding 40–50 million reads per library.