Regensburg 2007 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 21: High-Throughput Data and their Analysis

BP 21.1: Vortrag

Mittwoch, 28. März 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.