Nanopore based sequence specific detection of duplex DNA for genomic profiling.
Nanopore based sequence specific detection of duplex DNA for genomic profiling.
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DOI：
10.1021/nl100058y
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发表时间：
2010-02-10
期刊：
影响因子：
10.8
通讯作者：
Meller A
中科院分区：
文献类型：
--
作者：
Singer A;Wanunu M;Morrison W;Kuhn H;Frank-Kamenetskii M;Meller A
我们展示了一种纯电学的单分子检测特定DNA序列的方法，通过将双链DNA(DsDNA)与肽核酸(PNA)探针杂交并通过亚5 nm氮化硅小孔电泳线连接DNA实现的。为了获得高亲和力和序列特异性，采用BIS-PNA作为标记探针。序列检测是通过读取单个移位DNA分子的离子电流轨迹来执行的，这些移位DNA分子显示出特征的次级阻断水平，而未标记的分子中没有这种水平。通过对同一基因组片段上不同位置的一次标记和两次标记的DNA进行纳米孔分析，证明了对DNA进行条形码的可能性。我们的高通量、长阅读长度的方法可以用来识别嵌入单个DNA分子的关键序列，而不需要扩增或荧光/放射性标记。这为人类基因组学以及抗击传染病的病原体检测开辟了广泛的可能性。
We demonstrate a purely electrical method for single-molecule detection of specific DNA sequences, achieved by hybridizing double-stranded DNA (dsDNA) with peptide nucleic acid (PNA) probes and electrophoretically threading the DNA through sub-5 nm silicon nitride pores. Bis-PNAs were used as the tagging probes, in order to achieve high affinity and sequence-specificity. Sequence detection is performed by reading the ion current traces of individual translocating DNA molecules, which display a characteristic secondary blockade level, absent in untagged molecules. The potential for barcoding DNA is demonstrated through nanopore analysis of once-tagged and twice-tagged DNA at different locations on the same genomic fragment. Our high-throughput, long-read length method can be used to identify key sequences embedded in individual DNA molecules, without the need for amplification or fluorescent/radio labeling. This opens up a wide range of possibilities in human genomics, as well as in pathogen detection for fighting infectious diseases.
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影响因子：
10.8
作者：
Wanunu M;Sutin J;Meller A
通讯作者：
Meller A
影响因子：
15
作者：
He, Gaofei;Rapireddy, Sirinivas;Ly, Danith H.
通讯作者：
Ly, Danith H.
影响因子：
3.4
作者：
Cherny, DI;Fourcade, A;Delain, E
通讯作者：
Delain, E
DOI：
10.1073/pnas.96.21.11804
发表时间：
1999-10-12
影响因子：
11.1
作者：
Lohse, J;Dahl, O;Nielsen, PE
通讯作者：
Nielsen, PE
影响因子：
3.4
作者：
Wanunu, Meni;Sutin, Jason;Meller, Amit
通讯作者：
Meller, Amit