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Figure 2
MS-HRM standard curves of STAT1. (A) Aligned melt curves, (B) Difference plot & (C) Derivative melt curves of 100%, 75%, 50%, 25%, 10% and 0% methylated DNA standards, normalized to the 0% methylated DNA. Aligned melt curves (A) show that the 100% methylated DNA had higher melting temperature as compared to unmethylated DNA for each gene analyzed by MS-HRM. The 75%, 50%, 25% and 10% methylated DNA standards had a melting temperature situated between the 100% and unmethylated DNA standards which is respective of the percentage methylation of sample. The graph of the negative first derivative of the melting curve (C) shows that the 100% methylated and unmethylated DNA had only one melt peak, whereas 75%, 50%, 25% and 10% standards had two melt peaks representative of each of the DNA standard.
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