DNA sequencing and bar-coding using solid-state nanopores.
DNA sequencing and bar-coding using solid-state nanopores.
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DOI：
10.1002/elps.201200266
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发表时间：
2012-12
期刊：
影响因子：
2.9
通讯作者：
Meller, Amit
中科院分区：
文献类型：
--
作者：
Atas, Evrim;Singer, Alon;Meller, Amit
纳米孔已经成为一种突出的单分子分析工具，特别有希望用于基因组应用。在这篇综述中，我们讨论了纳米孔传感器的两个潜在应用：首先，我们提出了一种基于纳米孔的单分子DNA测序方法，该方法利用光学检测进行大规模并行处理。其次，我们描述了一种方法，通过该方法，纳米孔可以用作单分子基因分型工具。对于DNA测序，四种类型的DNA核碱基之间的区别是通过采用DNA扩增的生物化学程序来实现的。在这种方法中，每条DNA链中的每个核碱基在保留核碱基序列的过程中被转化为四个预定义的独特16聚体之一。然后将所得的转化链与四个分子信标的文库杂交，每个分子信标携带独特的荧光团标签，其是用于转化的16聚体的完美互补物。然后使用固态纳米孔依次去除这些信标，一个接一个，导致一系列可以被光学检测到的四种颜色的光子爆发。单分子基因分型通过用γ-修饰的合成肽核酸探针标记DNA片段来实现，所述探针使用固态纳米孔偶联到复合物的电子表征。该方法可用于鉴定和区分在单分子水平上具有高水平序列相似性但病理或对治疗的反应不同的基因。我们将通过区分两种高度相似的人类免疫缺陷病毒亚型的pol基因来说明这种方法，为病毒分类的新型诊断平台铺平道路。
Nanopores have emerged as a prominent single-molecule analytic tool with particular promise for genomic applications. In this review, we discuss two potential applications of the nanopore sensors: First, we present a nanopore-based single-molecule DNA sequencing method that utilizes optical detection for massively parallel throughput. Second, we describe a method by which nanopores can be used as single-molecule genotyping tools. For DNA sequencing, the distinction among the four types of DNA nucleobases is achieved by employing a biochemical procedure for DNA expansion. In this approach, each nucleobase in each DNA strand is converted into one of four predefined unique 16-mers in a process that preserves the nucleobase sequence. The resulting converted strands are then hybridized to a library of four molecular beacons, each carrying a unique fluorophore tag, that are perfect complements to the 16-mers used for conversion. Solid-state nanopores are then used to sequentially remove these beacons, one after the other, leading to a series of photon bursts in four colors that can be optically detected. Single-molecule genotyping is achieved by tagging the DNA fragments with γ-modified synthetic peptide nucleic acid probes coupled to an electronic characterization of the complexes using solid-state nanopores. This method can be used to identify and differentiate genes with a high level of sequence similarity at the single-molecule level, but different pathology or response to treatment. We will illustrate this method by differentiating the pol gene for two highly similar human immunodeficiency virus subtypes, paving the way for a novel diagnostics platform for viral classification.
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影响因子：
10.8
作者：
McNally B;Wanunu M;Meller A
通讯作者：
Meller A
DOI：
10.1002/cbic.200800441
发表时间：
2008-10-13
期刊：
Chembiochem : a European journal of chemical biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
10.8
作者：
McNally B;Singer A;Yu Z;Sun Y;Weng Z;Meller A
通讯作者：
Meller A
影响因子：
10.8
作者：
Nam, Sung-Wook;Rooks, Michael J.;Rossnagel, Stephen M.
通讯作者：
Rossnagel, Stephen M.
影响因子：
15
作者：
He, Gaofei;Rapireddy, Sirinivas;Ly, Danith H.
通讯作者：
Ly, Danith H.