DNA methylation profiling Whole genome profiling - human peripheral blood mononuclear cells

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

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Quantification of nuclear DNMT3A and DNMT3B enzyme concentrations were performed as per the manufacturer's guidelines (Epiquik DNMT3A/B Assay Kit, Epigentek, New York). All samples were assayed in duplicate, and various dilutions of the respective DNMT standards were used to plot a standard curve. Microplates were scanned at 450 nm with a reference wavelength of 655 nm. Intra‐assay coefficient of variations for DNMT3A and DNMT3B were calculated as 3.95% and 9.92%, respectively.
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