Chemically Tuned Silicon Nitride Nanopores for Nucleic Acid Sequencing

项目摘要 该项目旨在将DNA和RNA测序技术至少提高一个数量级， 通过显著提高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。虽然氮化硅是传统的 用于纳米孔测序应用的材料，其复杂的带电天然表面化学 对于带电核酸生物聚合物来说， 穿过比它本身大不了多少的纳米孔。非常理想的长读取长度 增加了对纳米孔表面的化学控制的需要。涂覆纳米孔 甚至具有单个分子层的表面将改变纳米孔直径，这因此也 提供了纳米孔尺寸的分子级调节。广泛地，化学调谐 纳米孔表面化学提供对（天然或标记的）核酸运动的控制 通过静电、特定的化学相互作用， 电动力学（例如电渗）。它提供了潜在的钝化污垢， 复杂的矩阵，从而支持更小的样品处理。它还提供了控制 界面现象，可以影响纳米孔电流噪声。 氮化硅薄膜是一种应用广泛的纳米纤维材料，具有广泛的商业应用前景。 实用性，使得其在许多纳米孔测序实现中的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在该领域中没有获得牵引力，这在很大程度上是因为 该化学品固有地具有实施挑战性，考虑到化学品的可变性， 氮氧化硅涂层。因此，我们建议开发一种完全不同类型的表面 化学改性策略易于实施，产生高度可靠的结果， 其可用于安装具有各种化学性质和尺寸的表面涂层。 我们建议通过它们对纳米孔电导的影响来测试表面涂层， 电流噪声，以及当感测明确定义的 DNA序列。 该项目将由一个跨学科的团队实施，该团队结合了20多年的经验， 主要研究者和合作研究者在物理有机化学和化学 合成（MK）;材料科学（MK&JRD）;和纳米纤维（JRD）， 在纳米孔科学的经验开始在纳米孔基因分型（JRD），包括专业化 纳米孔表面化学修饰和表征。

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.