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Once samples were ready, RT-qPCR was performed to detect the mRNA expression levels of miR-27a-3p and miR-27b-3p using 2 l of cDNA that was added to 10 l of PCR mix prepared by adding 0.5 l of TaqMan MicroRNA assay 20X, 5 l PCR master mix (TaqMan Fast Advanced Master mix, #4444964, Applied Biosystem), and 4.5 l of sterile water