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Thermal denaturation studies provide a simple and unambiguous demonstration of drug binding to DNA. Small molecules that bind preferentially to the DNA duplex stabilize the structure and elevate its melting temperature (Tm). Apart from a simple qualitative demonstration of binding to DNA, melting studies can be analysed to obtain quantitative information about the binding interaction. One purpose of this chapter is to illustrate the utility of such methods in the quantitative analysis of drug–DNA melting curves to obtain thermodynamic data. A powerful high-throughput screening method for the discovery of nucleic acid binders, differential scanning fluorometry (DSF), will be described.

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