GSM9886122: JJ102_Mettl3_KO; Mus musculus; RNA-Seq
Mouse esophageal epithelial cells were prepared as single-cell suspensions and enriched for EpCAM⁺ epithelial cells while excluding dead cells and immune cells by FACS. Four biological replicates per group were included for bulk RNA-seq analysis. Total RNA was extracted using the RNeasy Plus Micro Kit, quantified with Qubit and NanoDrop, and quality-checked using a Bioanalyzer. Samples with RIN ≥ 8 were used for library preparation. Poly(A)⁺ mRNA was enriched, and strand-specific libraries were prepared using the NEBNext Ultra II kit with an insert size of 200–300 bp. Libraries were sequenced on an Illumina NovaSeq 6000 platform with paired-end 150-bp reads, generating ≥40 million read pairs per sample, with Q30 ≥ 85%.