AVS 52nd International Symposium
    DNA Topical Conference Monday Sessions
       Session DN-MoP

Paper DN-MoP6
SPR Imaging Detection of DNA Hybridization/Adsorption with Exonuclease III Enzymatic Reaction

Monday, October 31, 2005, 5:00 pm, Room Exhibit Hall C&D

Session: DNA Poster Session
Presenter: H.J. Lee, University of California, Irvine
Authors: H.J. Lee, University of California, Irvine
R.M. Corn, University of California, Irvine
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DNA microarrays are emerging as a powerful tool for gene analysis, viral identification, medical diagnostics and many other biological applications. The next generation of DNA microarrays for biosensing will utilize both surface bioaffinity interactions and subsequent surface enzyme reactions on the adsorbed species in order to greatly enhance the specificity and sensitivity of the biosensor. For instance, we have recently demonstrated a novel approach that utilizes the enzyme RNase H in conjunction with RNA microarrays for the direct detection of DNA at femtomolar concentrations with surface plasmon resonance (SPR) imaging. In this presentation, we demonstrate the use of enzyme Exonuclease III, which specifically digests duplex DNA, in conjunction with DNA microarrays for the amplified SPR imaging detection of single stranded target DNA. Kinetic measurements of the surface Exo III reaction as a function of temperature and enzyme concentration were performed using a combination of SPR imaging and SPR fluorescence. Greater insight into the surface enzyme reaction was achieved through analysis using a newly developed kinetics model that couples the contributions of both enzyme adsorption and surface enzyme reaction kinetics.