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BP: Fachverband Biologische Physik
BP 21: High-Throughput Data and their Analysis
BP 21.1: Talk
Wednesday, March 28, 2007, 18:15–18:30, H43
Competitive DNA Hybridization: Experimental Results and Conclusions for Microarray Experiments — •Timo Mai1, Wolfgang Michel1, Philipp Baaske2, Thomas Naiser1, and Albrecht Ott1 — 1Physikalisches Institut, Universität Bayreuth, 95440 Bayreuth — 2present address: Lehrstuhl für Experimentelle Physik - Biophysik, LMU München, 80539 München
We experimentally approach the complex multi-component hybridization to microarrays by a simple system: A two-component mixture in solution consisting of a perfect match (PM) and a mismatch (MM) oligonucleotide competing for a immobilized probe sequence. We first characterize the binding of PM and MM separately in single hybridization experiments. We deduce rate constants and confirm them with values extracted from analysis of the temperature dependent hybridization signal. From this we predict the time course of hybridization of PM and MM when in competition for surface binding sites and compare with our competitive hybridization experiments.
This is useful for understanding the process of MM displacement by the PM and attaining an estimate for the specificity and the detection limit of microarray experiments.