AVS 53rd International Symposium
    Nucleic Acids at Surfaces Topical Conference Monday Sessions
       Session DN-MoA

Paper DN-MoA9
Achieving Reliable Microarray Analysis Results Using Competitive Hybridization

Monday, November 13, 2006, 4:40 pm, Room 2014

Session: Nucleic Acids at Surfaces II
Presenter: A. Chagovetz, University of Utah
Authors: A. Chagovetz, University of Utah
L. Williams, University of Utah
J. Bishop, University of Utah
S. Blair, University of Utah
Correspondent: Click to Email
As microarrays migrate from detection of dissimilar analytes to the detection of analytes that have only a single to a few base mismatches, surface hybridization may become competitive. We have simulated competitive hybridization using a finite element analysis model. Our results show that observed dynamic range between the complement and its competitors increases with temperature because of enhanced dissociation of the mismatch target, assuming thermodynamic equilibrium is not reached. Additionally, competitive hybridization can be enhanced by decreasing the immobilized probe concentration on the surface of the substrate. Using the two observations above we propose two different analysis methods for hybridization experiments, one which applies to end-point analysis and the other which is for real-time analysis. The first method uses a labeled multianalyte sample, while the second method is a novel label-less detection mechanism for multianalyte samples. By introducing a label target that is known to be of a lower affinity than the targets to be investigated we can watch the dissociation of the lower affinity species and predict the concentration of the complement. This is verified experimentally using a microscale array of individually controlled heating elements. The heater array was developed at Sandia National Laboratories, using surface micromachining technology.