Cantilever-based optical deflection assay for discrimination of DNA single-nucleotide mismatches

K. M. Hansen, H. F. Ji, G. Wu, R. Datar, R. Cote, A. Majumdar, T. Thundat

Research output: Contribution to journalArticle

324 Scopus citations

Abstract

Characterization of single-nucleotide polymorphisms is a major focus of current genomics research. We demonstrate the discrimination of DNA mismatches using an elegantly simple microcantilever-based optical deflection assay, without the need for external labeling. Gold-coated silicon AFM cantilevers were functionalized with thiolated 20- or 25-mer probe DNA oligonucleotides and exposed to target oligonucleotides of varying sequence in static and flow conditions. Hybridization of 10-mer complementary target oligonucleotides resulted in net positive deflection, while hybridization with targets containing one or two internal mismatches resulted in net negative deflection. Mismatched targets produced a stable and measurable signal when only a four-base pair stretch was complementary to the probe sequence. This technique is readily adaptable to a high-throughput array format and provides a distinct positive/negative signal for easy interpretation of oligonucleotide hybridization.

Original languageEnglish (US)
Pages (from-to)1567-1571
Number of pages5
JournalAnalytical Chemistry
Volume73
Issue number7
DOIs
StatePublished - Apr 1 2001

ASJC Scopus subject areas

  • Analytical Chemistry

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