Sequence-specific Nucleic Acid Detection from Binary Pore Conductance Measurement
Sequence-specific Nucleic Acid Detection from Binary Pore Conductance Measurement
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DOI：
10.1021/ja3059205
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发表时间：
2012-09-26
影响因子：
15
通讯作者：
Schmidt, Jacob J.
中科院分区：
文献类型：
--
作者：
Esfandiari, Leyla;Monbouquette, Harold G.;Schmidt, Jacob J.
我们描述了一种用于序列特异性核酸(NA)检测的平台，利用直径为2微米的微管和直径为3微米的聚苯乙烯小球，未带电的肽核酸(PNA)探针分子已连接到该微管上。当目标NAS与互补的PNA微珠杂交时，微珠获得负电荷并变得可电泳动。外加电场引导这些NA-PNA微珠朝向吸管尖端，它们被阻挡，导致无限期的、电可检测的、部分堵塞毛孔。在非互补NA的存在下，即使达到单碱基错配的水平，也看不到永久性的孔堵塞。我们展示了该平台在炭疽致死因子序列检测中的应用。
We describe a platform for sequence-specific nucleic acid (NA) detection utilizing a micropipet tapered to a 2 mu m diameter pore and 3 mu m 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.