2.8. ELISA‐based global genomic DNA methylation quantification

Global DNA methylation was quantified as the percentage relative to control methylated DNA by using the anti‐5mC antibody from Imprint^@ Methylated DNA Quantification Kit as per manufactures instructions manual supplied with the kit (Sigma–Aldrich; Catalog number—MDQ1). Three biological and three technical replicates for each sample were used. The environmental error which may occur without enough replicates was normalized. To calculate methylation levels relative to the control methylated DNA sample, the following formula was applied: [(A _{450 av} Sample − A _{450 av} Blank)/(A _{450 av} Methylated Control DNA − A _{450 av} Blank)] × 100 where the A450 av represent the averages of the blank, the sample and the methylated control DNA replicates.