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Originally published as Biophys J. BioFAST on November 3, 2006.
doi:10.1529/biophysj.106.097790
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Biophysical Journal 92:935-946 (2007)
© 2007 The Biophysical Society

Temperature Effects on DNA Chip Experiments from Surface Plasmon Resonance Imaging: Isotherms and Melting Curves

J. B. Fiche, A. Buhot, R. Calemczuk and T. Livache

SPrAM (Structure et Propriétés d'Architectures Moléculaires) UMR 5819 (Commissariat à l'Énergie Atomique, Centre Nationale de Recherche Scientifique, Université Joseph Fourier), Département de Recherche Fondamentale sur la Matière Condensée, CEA Grenoble, France

Correspondence: Address reprint requests to Arnaud Buhot, SPrAM UMR 5819, DRFMC, CEA Grenoble, 17 rue des Martyrs, 38054 Grenoble cedex 9, France. E-mail: arnaud.buhot{at}cea.fr.

We present an analysis of hybridization experiments on a DNA chip studied by surface plasmon resonance imaging. The reaction constants at various temperatures and for different probe lengths are obtained from Langmuir isotherms and hybridization kinetics. The melting curves from temperature scans are also obtained without any labeling of the targets. The effects of the probe length on the hybridization thermodynamics, deduced from the temperature dependence of the reaction constants as well as from the melting curves, suggest dispersion in the length of the hybridization segments of the probes accessible to the targets. Those are, however, sufficient to suggest efficient point mutation detection from temperature scans.




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