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To quantify mRNA, following cDNA synthesis, the quantitative RT-PCR (qRT-PCR) analysis was carried out on an Applied Rotor-Gene 3000 with the following conditions: GAPDH and target genes were amplified for 40 cycles at 94C for 30 seconds, 63C for 60 seconds, and 72C for 90 seconds

These changes in methylation are early events in tumorigenesis

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Comparing Western-type diet (WD) and fiber-enriched Mediterranean diet (FMD), the abundance of Firmicutes decreased, Bacteroidetes and Proteobacteria levels increased, and SCFA content increased, especially propionate and butyrate after FMD (Soldn et al., 2024)