Label-free "digital detection" of single-molecule DNA hybridization with a single electron transistor
Label-free "digital detection" of single-molecule DNA hybridization with a single electron transistor
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DOI：
10.1021/ja063022f
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发表时间：
2006-09-06
影响因子：
15
通讯作者：
Brousseau, Louis C., III
中科院分区：
文献类型：
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作者：
Brousseau, Louis C., III
在这里，我们提出了一种新的分析方法，消除了荧光标记，使单分子DNA杂交在复杂基质中的“数字检测”具有极大的简化方案。单个寡核苷酸的结合态与单电子晶体管（SET）的量子点（QD）的电子耦合通过“分子门控”提供了对结合事件的实时直接观察。与分子捕获相关的静电电荷的变化被用来代替栅极来调节SET电导率。含有碱基错配的目标寡核苷酸在室温下0.1X SSC下不会引起SET反应，离子强度或ph值也不会发生变化。此外，即使在光学不可接近的基质中，如血清或硫氰酸铵裂解缓冲液中也可以检测到杂交。
Here we present a novel assay that eliminates fluorescent labels and enables “digital detection” of single-molecule DNA hybridization in complex matrixes with greatly simplified protocols. Electronic coupling of the binding state of a single oligonucleotide to the quantum dot (QD) of a single electron transistor (SET) affords direct observation of binding events in real-time via “molecular gating”. The change of electrostatic charge associated with the molecular capture is used in lieu of a gate electrode to modulate the SET conductivity. Target oligos containing base mismatches do not elicit SET response under 0.1X SSC at room temperature nor do changes in ionic strength or pH. Furthermore, hybridization is detected even in optically inaccessible matrixes such as serum or quanidinium thiocyanate lysis buffer.