GSM9558064: Mouse podocyte USP40 siRNA#1 biol rep 1; Mus musculus; RNA-Seq
Total RNA was extracted and RNA quality was assessed using the Agilent RNA 6000 Nano kit (Cat#5067-1511) on a Bioanalyzer. One hundred nanograms of total RNA were evaluated for quality control, and 1250 ng of high-quality RNA were used for library preparation. Poly(A)+ RNA was enriched using the NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat#E7490). Strand-specific libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina (Cat#E7770L). Libraries were amplified with 12 PCR cycles. Library size distribution was validated using the Agilent DNA 7500 kit (Cat#5067-1506). Each biological replicate corresponds to an independent RNA-seq library. Paired-end sequencing (2 × 36 bp) was performed on an Illumina NextSeq 500 using the NextSeq 500/550 High Output Kit v2.5 (75 Cycles; Cat#20024906), generating two FASTQ files per sample (R1 and R2). Raw sequencing reads were adapter-trimmed and quality-filtered using CLC Genomics Workbench version 23.02.