A single-molecule barcoding system using nanoslits for DNA analysis
A single-molecule barcoding system using nanoslits for DNA analysis
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DOI：
10.1073/pnas.0611151104
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发表时间：
2007-02-20
影响因子：
11.1
通讯作者：
Schwartz, David C.
中科院分区：
文献类型：
--
作者：
Jo, Kyubong;Dhingra, Dalia M.;Schwartz, David C.
分子约束为排列大DNA分子提供了新的途径，使单分子方案能够获得序列信息。如果一个新生系统的元素能够在早期发展阶段被整合，这样的计划可以迅速发展成为能够进行基因组分析的平台。为了克服纳米级器件在制造、样品装载、生化标记和检测方面的严格实验要求，需要集成策略。我们证明，当缓冲条件被控制以改变DNA硬度时，具有微纳米级特征的一次性装置可以大大延长DNA分子。此外，我们提出了描述这种延伸的分析计算。我们还开发了一种互补的酶标记方案，该方案通过荧光共振能量转移成像，在所描述的纳米狭缝设备中标记细长分子上的特定序列。总的来说，这些发展为基因组分析提供了可扩展的分子限制方法。
Molecular confinement offers new routes for arraying large DNA molecules, enabling single-molecule schemes aimed at the acquisition of sequence information. Such schemes can rapidly advance to become platforms capable of genome analysis if elements of a nascent system can be integrated at an early stage of development. Integrated strategies are needed for surmounting the stringent experimental requirements of nanoscale devices regarding fabrication, sample loading, biochemical labeling, and detection. We demonstrate that disposable devices featuring both micro- and nanoscale features can greatly elongate DNA molecules when buffer conditions are controlled to alter DNA stiffness. Furthermore, we present analytical calculations that describe this elongation. We also developed a complementary enzymatic labeling scheme that tags specific sequences on elongated molecules within described nanoslit devices that are imaged via fluorescence resonance energy transfer. Collectively, these developments enable scaleable molecular confinement approaches for genome analysis.