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

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

Research output: Contribution to journalArticle

316 Citations (Scopus)

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

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Oligonucleotides
Assays
Nucleotides
DNA
DNA Probes
Silicon
Polymorphism
Gold
Labeling
Throughput

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Cantilever-based optical deflection assay for discrimination of DNA single-nucleotide mismatches. / Hansen, K. M.; Ji, H. F.; Wu, G.; Datar, Ram; Cote, Richard J; Majumdar, A.; Thundat, T.

In: Analytical Chemistry, Vol. 73, No. 7, 01.04.2001, p. 1567-1571.

Research output: Contribution to journalArticle

Hansen, K. M. ; Ji, H. F. ; Wu, G. ; Datar, Ram ; Cote, Richard J ; Majumdar, A. ; Thundat, T. / Cantilever-based optical deflection assay for discrimination of DNA single-nucleotide mismatches. In: Analytical Chemistry. 2001 ; Vol. 73, No. 7. pp. 1567-1571.
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