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Physicochemical study of DNA biochips: DNA duplex stability, point mutation detection and beyond

Abstract : This Ph.D. thesis studies phenomena concerning DNA duplex stability. Exploiting a real-time detection system based on Surface Plasmon Resonance imaging (SPRi), we use a temperature scan method to analyze DNA interactions on functionalized biochips. First, a detailed study about a comparison of two DNA immobilization methods and the influence of different buffer components on surface hybridized DNA duplexes is presented. Especially the influence of salt and denaturing agents in the buffer are discussed. Second, we apply the temperature scan method to point mutation detection, as well for oligonucleotides as for longer DNA or RNA targets produced by Polymerase Chain Reaction (PCR) or Nucleic Acid Sequence Based Amplication (NASBA) amplification protocols, respectively. Competition between targets containing point mutations and surface hybridization is addressed. Also, secondary structures in solution may alter target capture on probes. For the isothermal NASBA amplification, we show that it is possible to make an integrated system with amplification and detection on the SPRi chip. Finally, a study of DNA lesions and repair enzymes is presented in collaboration with the 'Laboratory of Nucleic Acids Lesions' (LAN) at the CEA Grenoble profiting once again from the flexible temperature regulation to characterize enzyme activity on surface grafted DNA.
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https://tel.archives-ouvertes.fr/tel-00433465
Contributor : Julia Fuchs <>
Submitted on : Thursday, November 19, 2009 - 2:30:09 PM
Last modification on : Wednesday, November 4, 2020 - 1:59:53 PM
Long-term archiving on: : Thursday, June 17, 2010 - 6:40:50 PM

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  • HAL Id : tel-00433465, version 1

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Julia Fuchs. Physicochemical study of DNA biochips: DNA duplex stability, point mutation detection and beyond. Biological Physics [physics.bio-ph]. Université Joseph-Fourier - Grenoble I, 2009. English. ⟨tel-00433465⟩

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