Chemically Tuned Silicon Nitride Nanopores for Nucleic Acid Sequencing

项目总结 该项目旨在将DNA和RNA测序技术提高至少一个数量级 极大地提高了控制氮化硅纳米孔表面的能力 化学和改性氮化硅纳米孔尺寸。而氮化硅是一种传统的 纳米孔测序材料及其络合物带电天然表面化学 会给带电的核酸生物聚合物带来一个具有挑战性的复杂环境 穿过一个比自己大不了多少的纳米孔。非常理想的长阅读长度 提高了对纳米孔表面进行化学控制的必要性。包覆纳米孔 即使只有一个分子层的表面也会改变纳米孔的直径，因此 提供了纳米孔尺寸的分子级调节。广泛地说，化学上的调整 纳米孔表面化学可以控制(天然的或标记的)核酸的运动 聚合物通过静电学、特定的化学相互作用通过孔，以及 电动力学(例如电渗法)。它提供了防止污垢的钝化潜力 复数矩阵，从而支持更小的样本处理。它还提供了控制 可能影响纳米孔电流噪声的界面现象。 薄膜氮化硅是一种用途广泛的纳米加工材料，具有广泛的商业应用前景 实用程序，使其在许多纳米孔测序实施中继续使用是有保证的 尽管它的表面化学经常具有挑战性。但控制和改善其表面的努力 使用硅烷化学的化学并没有在该领域获得吸引力，这在很大程度上是因为 化学的实现本身就具有挑战性，考虑到 氮化硅氧化物涂层。因此，我们建议开发一种完全不同类型的曲面 易于实施的化学修饰策略，产生高度可靠的结果，以及 可用于安装具有各种化学性质和尺寸的表面涂层。 我们建议通过测试表面涂层对纳米孔电导和 电流噪声，以及在检测时特定于序列的信号特征 DNA序列。 该项目将由一个结合了20多年的跨学科团队来实施 在物理、有机化学和化学方面的主要和合作研究人员的经验 合成(MK)；材料科学(MK&JRD)；纳米制造(JRD)，十年 从纳米孔基因分型(JRD)开始的纳米孔科学经验，包括专业知识 在纳米孔表面化学修饰和表征方面。

PROJECT SUMMARY This project aims to improve DNA and RNA sequencing technology by at least an order of magnitude by dramatically improving the ability to control silicon nitride nanopore surface chemistry and to modify silicon nitride nanopore size. While silicon nitride is a conventional material for nanopore sequencing applications, its complex charged native surface chemistry can present a challenging and complicated environment for a charged nucleic acid biopolymer passing through a nanopore not much larger than itself. Highly desirable long read lengths heighten the need for chemical control over the nanopore surface. Coating the nanopore surface with even a single molecular layer will change the nanopore diameter, which thus also provides for molecular-scale tuning of nanopore dimensions. Broadly, chemically tuned nanopore surface chemistry affords control over motion of (native or labelled) nucleic acid polymers through the pore through electrostatics, specific chemical interactions, and electrokinetics (e.g. electroosmosis). It offers the potential for passivation against fouling in complex matrices, thereby supporting more minimal sample processing. It also affords control over interfacial phenomena that can affect nanopore current noise. Thin-film silicon nitride is a widely used nanofabrication material with widespread commercial utility, so that its continued use in a host of nanopore sequencing implementations is warranted in spite of its often challenging surface chemistry. But efforts to control and improve its surface chemistry using silane chemistry have not gained traction in the field, in significant part because the chemistry is inherently challenging to implement, the more so given the variability of the silicon nitride oxide coating. We thus propose to develop a radically different type of surface chemical modification strategy that is simple to implement, produces highly reliable results, and that can be used to install surface coatings with a wide variety of chemical properties and sizes. We propose to test the surface coatings through their effect on the nanopore conductance and current noise, and on the sequence-specific signal characteristics when sensing well-defined sequences of DNA. The project will be implemented by an interdisciplinary team that combines more than 20 years of Principal and Co-Investigator experience in physical organic chemistry and chemical synthesis (MK); materials science (MK&JRD); and nanofabrication (JRD), with a decade of experience in nanopore science begun in nanopore genotyping (JRD), including a specialization in nanopore surface chemistry modification and characterization.