RNA-seq of Streptococcus mutans
Total RNA was extracted using TRIzol reagent, and RNA quality was assessed prior to library construction. mRNA was enriched using poly(T) magnetic beads, fragmented, and reverse-transcribed to generate cDNA libraries, which were subsequently end-repaired, adapter-ligated, PCR-amplified, and sequenced on an Illumina NovaSeq 6000 platform. Raw sequencing reads were quality-filtered using fastp, and clean reads were aligned to the reference genome with HISAT2. Gene expression levels were quantified using HTSeq and normalized as FPKM values. Differentially expressed genes were identified using DESeq with |logFoldChange| > 1 and adjusted P < 0.08. Functional enrichment analyses, including Gene Ontology and KEGG pathway analyses, were performed using clusterProfiler, and gene set enrichment analysis was conducted using GSEA software.
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