GSM9900929: E2-30-7; Mus musculus; RNA-Seq
Total RNA was extracted from mouse renal tissue using TRIzol reagent (Invitrogen) following the manufacturer's instructions. RNA purity and concentration were determined by Nanodrop 2000 spectrophotometer. RNA integrity was verified via agarose gel electrophoresis and Agilent 2100 Bioanalyzer; only samples with RIN ≥ 7.0 were retained for library construction. mRNA was enriched from total RNA using oligo(dT) magnetic beads, fragmented into 300–350 bp fragments. First-strand and second-strand cDNA were synthesized, followed by end repair, A-tailing and sequencing adapter ligation. Purified cDNA fragments were amplified by PCR to construct RNA-seq libraries. Qualified libraries were sequenced on Illumina NovaSeq 6000 platform with paired-end 150 bp sequencing strategy.