Analysis of Multidimensional G‐Quadruplex Melting Curves

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Abstract
Multidimensional “3D” melting curves for G‐quadruplexes are obtained by recording whole spectra (absorbance, CD, fluorescence) as a function of temperature, rather than the common approach of recording the spectral response to temperature at a single wavelength. 3D melting curves are richer in information, and can be used to enumerate the number of significant species and intermediate states required to properly analyze the thermal denaturation reaction to obtain thermodynamic information. This unit describes the application of the method of singular value decomposition to the analysis of 3D melting data obtained for G‐quadruplex structures, and how the results of such an analysis can be used to provide a more complete characterization of the mechanism of quadruplex unfolding. Curr. Protoc. Nucleic Acid Chem. 45:17.4.1‐17.4.16. © 2011 by John Wiley & Sons, Inc.