Single-molecule electrical random resequencing of DNA and RNA.
Single-molecule electrical random resequencing of DNA and RNA.
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DOI：
10.1038/srep00501
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发表时间：
2012
影响因子：
4.6
通讯作者：
Kawai, Tomoji
中科院分区：
文献类型：
--
作者：
Ohshiro, Takahito;Matsubara, Kazuki;Tsutsui, Makusu;Furuhashi, Masayuki;Taniguchi, Masateru;Kawai, Tomoji
DNA测序技术的两个范式转变-从批量到单分子和从光学到电学检测-有望实现不需要PCR扩增的无标记、低成本DNA测序。它将导致个性化医疗的高通量第三代测序技术的发展。虽然纳米孔装置已被提议作为第三代DNA测序装置，但通过展示使用电信号对DNA进行重新测序的新技术，已经实现了这些技术中的重要里程碑。在这里，我们报告的单分子电重测序的DNA和RNA使用的混合方法，通过隧道电流和随机测序识别单碱基分子。我们的方法读取九种类型的DNA寡聚体的序列。来自let-7 microRNA家族的5′-UGAGGUA-3′的完整序列也通过创建重叠片段序列的复合物来鉴定，该复合物是使用单碱基分子在一对纳米电极之间通过时传导的隧道电流随机确定的。
Two paradigm shifts in DNA sequencing technologies—from bulk to single molecules and from optical to electrical detection—are expected to realize label-free, low-cost DNA sequencing that does not require PCR amplification. It will lead to development of high-throughput third-generation sequencing technologies for personalized medicine. Although nanopore devices have been proposed as third-generation DNA-sequencing devices, a significant milestone in these technologies has been attained by demonstrating a novel technique for resequencing DNA using electrical signals. Here we report single-molecule electrical resequencing of DNA and RNA using a hybrid method of identifying single-base molecules via tunneling currents and random sequencing. Our method reads sequences of nine types of DNA oligomers. The complete sequence of 5′-UGAGGUA-3′ from the let-7 microRNA family was also identified by creating a composite of overlapping fragment sequences, which was randomly determined using tunneling current conducted by single-base molecules as they passed between a pair of nanoelectrodes.
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