Rapid Identification of DNA Fragments through Direct Sequencing with Electro-Optical Zero-Mode Waveguides.
Rapid Identification of DNA Fragments through Direct Sequencing with Electro-Optical Zero-Mode Waveguides.
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通过电光零模波导直接测序快速识别 DNA 片段。
DOI：
10.1002/adma.202209376
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Wanunu,Meni
中科院分区：
文献类型：
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作者：
Farhangdoust,Fatemeh;Alibakhshi,MohammadAmin;Cheng,Feng;Liang,Wentao;Liu,Yongmin;Wanunu,Meni
与序列特异性技术如聚合酶链反应（PCR）相比，DNA测序不需要事先了解样品的DNA。然而，目前的测序技术需要高输入用于合适的文库制备，这通常需要DNA扩增，即使对于单分子测序方法也是如此。在这里，我们提出了电光零模波导（eZMWs），它可以加载DNA到零模波导的限制，具有高效率和可忽略的DNA片段长度偏差。使用eZMW，我们观察到从超低输入（ng至pg水平）的各种大小的DNA片段的高效电压诱导的加载。我们展示了快速的DNA片段鉴定的短和长的DNA分子（260 bp和20，000 bp）的突发测序加载从一个等摩尔浓度的混合物在短短几分钟内。我们的设备允许进一步的研究，其中低输入DNA捕获是必不可少的，例如，在表观遗传学中，需要天然DNA来获得修饰的碱基信息。
In contrast to sequence-specific techniques such as polymerase chain reaction (PCR), DNA sequencing does not require prior knowledge of the sample for surveying DNA. However, current sequencing technologies demand high inputs for a suitable library preparation, which typically necessitates DNA amplification, even for single-molecule sequencing methods. Here, we present electro-optical zero-mode waveguides (eZMWs), which can load DNA into the confinement of zero-mode waveguides with high efficiency and negligible DNA fragment length bias. Using eZMWs we observe highly efficient voltage-induced loading of DNA fragments of various sizes from ultralow inputs (ng-to-pg levels). We demonstrate rapid DNA fragment identification by burst sequencing of short and long DNA molecules (260 bp and 20,000 bp) loaded from an equimolar pM-level concentration mixture in just a few-minutes. Our device allows further studies in which low-input DNA capture is essential, for example, in epigenetics, where native DNA is required for obtaining modified base information.