Cantilever-based optical deflection assay for discrimination of DNA single-nucleotide mismatches
Cantilever-based optical deflection assay for discrimination of DNA single-nucleotide mismatches
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DOI：
10.1021/ac0012748
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发表时间：
2001-04-01
影响因子：
7.4
通讯作者：
Thundat, T
中科院分区：
文献类型：
--
作者：
Hansen, KM;Ji, HF;Thundat, T
单核苷酸多态性的表征是当前基因组学研究的主要焦点。我们证明了使用一个优雅简单的基于微电子显微镜的光学偏转分析，而不需要外部标记的DNA错配的歧视。金包被的硅原子力显微镜杠杆官能化与硫醇化的20-或25-mer探针DNA寡核苷酸，并暴露于不同序列的目标寡核苷酸在静态和现在的条件。10-mer互补靶寡核苷酸的杂交导致净正偏转，而与含有一个或两个内部错配的靶的杂交导致净负偏转。错配的目标产生一个稳定的和可测量的信号时，只有四个碱基对的延伸是互补的探针序列。该技术易于适用于高通量阵列格式，并提供独特的阳性/阴性信号，便于解释寡核苷酸杂交。
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.