4.5 Article

Visual, base-specific detection of nucleic acid hybridization using polymerization-based amplification

Journal

ANALYTICAL BIOCHEMISTRY
Volume 386, Issue 2, Pages 285-287

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2008.12.009

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Funding

  1. State of Colorado
  2. University of Colorado Technology Transfer Office
  3. National Institutes of Health [R21 CA 127884]
  4. US Department of Education
  5. Teets Family Endowed Doctoral Fellowship in Nanotechnology

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Polymerization-based signal amplification offers sensitive visualization of biotinylated biomolecules functionalized to glass microarrays in a manner suitable for point-of-care use. Here we report using this method for visual detection of multiplexed nucleic acid hybridizations from complex media and develop an application toward point Mutation detection and single nucleotide polymorphism (SNP) typing, Primer extension reactions were employed to label selectively and universally all complementary surface DNA hybrids with photoinitiators, permitting simultaneous and dynamic photopolymerization from positive sites to 0.5-nM target concentrations. Dramatic improvements in signal ratios between complementary and mismatched hybrids enabled visual discrimination of single base differences in KRAS codon-12 biomarkers. (C) 2008 Elsevier Inc. All rights reserved.

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