从二元孔导测量中进行序列特异性核酸检测。
Sequence-specific nucleic acid detection from binary pore conductance measurement.
机构信息
Department of Bioengineering, University of California, Los Angeles, California, United States.
出版信息
J Am Chem Soc. 2012 Sep 26;134(38):15880-6. doi: 10.1021/ja3059205. Epub 2012 Sep 12.
Abstract
We describe a platform for sequence-specific nucleic acid (NA) detection utilizing a micropipet tapered to a 2 μm diameter pore and 3 μm diameter polystyrene beads to which uncharged peptide nucleic acid (PNA) probe molecules have been conjugated. As the target NAs hybridize to the complementary PNA-beads, the beads acquire negative charge and become electrophoretically mobile. An applied electric field guides these NA-PNA-beads toward the pipet tip, which they obstruct, leading to an indefinite, electrically detectable, partial blockade of the pore. In the presence of noncomplementary NA, even to the level of single base mismatch, permanent pore blockade is not seen. We show application of this platform to detection of the anthrax lethal factor sequence.
摘要
我们描述了一种利用微吸管（缩至 2 μm 直径的孔和 3 μm 直径的聚苯乙烯珠）的平台进行序列特异性核酸（NA）检测，其中未带电的肽核酸（PNA）探针分子已连接到珠上。当靶 NA 与互补的 PNA-珠杂交时，珠获得负电荷并变得电泳移动。施加的电场将这些 NA-PNA-珠引导到吸管尖端，它们会阻塞尖端，导致孔的无限电可检测的部分阻塞。在非互补 NA 的存在下，即使是单碱基错配的水平，也不会看到永久的孔阻塞。我们展示了该平台在炭疽致死因子序列检测中的应用。
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2
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3
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