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Equilibrium dissociation constant (KD) measurement via Isothermal titration calorimetry (ITC). The fully-, top- and bottom-methylated 54nt fragment with a central CmCGG pattern were performed as substrates for KD measurement via ITC. (A) KD for fully-methylated 54nt is 4.13 μM, and the binding molar ratio is ∼1.09. (B) KD for top-methylated 54nt is 3.14 μM, and the binding molar ratio is ∼0.71. (C) KD for bottom-methylated 54nt is 1.47 μM, and the binding molar ratio is ∼1.19.
