DNA methylation profiling Whole genome profiling - MCF-7, MDA-MB-453 human breast cancer

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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Protocol tips
gDNA was extracted using the NucleoSpin® Tissue kit (740952, Macherey Nagel). Global DNA methylation was quantified using the MethylFlash methylated DNA quantification kit (P-1034, Epigentek, France). Methyl DNA collection were performed using the “Methyl-Collector Ultra kit” (55005, Active Motif). Histone 3 acetylation was quantified using the EpiQuik Tissue Acetyl-Histone H3 ChIP Kit (P-2012, Epigentek). ChIP was performed using the ChIP-IT High Sensitivity kit (53040, Active Motif) with ChIP grade antibodies (Table ). All these kits were used according to the manufacturer’s instructions.
Protocol tips
gDNA was extracted using the NucleoSpin® Tissue kit (740952, Macherey Nagel). Global DNA methylation was quantified using the MethylFlash methylated DNA quantification kit (P-1034, Epigentek, France). Methyl DNA collection were performed using the “Methyl-Collector Ultra kit” (55005, Active Motif). Histone 3 acetylation was quantified using the EpiQuik Tissue Acetyl-Histone H3 ChIP Kit (P-2012, Epigentek). ChIP was performed using the ChIP-IT High Sensitivity kit (53040, Active Motif) with ChIP grade antibodies (Table ). All these kits were used according to the manufacturer’s instructions.
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