GSM9474937: F, Esr2KO, PBS, Rep 5; Mus musculus; RNA-Seq
Total RNA was extracted from snap-frozen whole lungs using the Zymo Quick-RNA Miniprep Kit (Zymo Research, Irvine, CA) following the manufacturer's protocol, including in-column DNase I treatment to remove genomic DNA. RNA purity and concentration were determined with a NanoDrop spectrophotometer (Thermo Fisher Scientific), and integrity was verified using an Agilent 2100 Bioanalyzer. Samples with RNA integrity number (RIN) ≥ 7.0 were used for sequencing. RNA-seq libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA) and sequenced on an Illumina NovaSeq 6000 at the Indiana University Center for Genomics and Bioinformatics (CGB) using paired-end 150 bp reads. Sequencing was conducted across two independent NovaSeq runs (CGB Project ID GSF3966), generating a total of 2.56 billion pass-filter reads (~30–40 million reads per sample).