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

被引:288
作者
Hansen, KM
Ji, HF
Wu, GH
Datar, R
Cote, R
Majumdar, A
Thundat, T
机构
[1] Oak Ridge Natl Lab, Div Life Sci, Oak Ridge, TN 37831 USA
[2] Univ Calif Berkeley, Dept Mech Engn, Berkeley, CA 94720 USA
[3] Univ So Calif, Dept Pathol, Los Angeles, CA 90033 USA
关键词
D O I
10.1021/ac0012748
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 now 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.
引用
收藏
页码:1567 / 1571
页数:5
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