Nanochannel-nanopore based DNA sequencing with DNA motion control and reduced entropic noise

基于纳米通道-纳米孔的 DNA 测序，具有 DNA 运动控制和降低的熵噪声

• 批准号: 10010924
• 负责人: David John Niedzwiecki
• 金额: $ 35万
• 依托单位: GOEPPERT, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-22 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10010924
• 关键词: Ablation; Address; Amplifiers; Area; Buffers; Caliber; Complex; Coupled; Custom; DNA; DNA sequencing; Detection; Development; Devices; Dimensions; Electrodes; Electrolytes; Electronics; Elements; Entropy; Enzymes; Event; Freedom; Genomics; Goals; Image; Ions; Label; Length; Measurement; Measures; Membrane; Methods; Microfluidics; Mission; Molecular Conformation; Monitor; Motion; National Human Genome Research Institute; Noise; Optics; Output; Performance; Phase; Photons; Polymerase; R43 grant; Running; Ships; Signal Transduction; Single-Stranded DNA; Small Business Innovation Research Grant; Speed; Stochastic Processes; Structure; System; Techniques; Technology; Temperature; Thinness; Time; United States National Institutes of Health; Variant; base; cost; feeding; fluorophore; improved; nanochannel; nanoimprint lithography; nanopore; new technology; novel; operation; sensor; sequencing platform; silicon nitride; single molecule; solid state; technology development; temporal measurement; transcriptome sequencing; voltage

Project Summary To improve DNA sequencing and to develop practical methods of RNA sequencing, this NHGRI Phase I project focuses on using solid-state nanopore sensors coupled with sub-30nm wide channels embedded into silicon nitride, with electronics operating at 10 MHz bandwidth for DNA sequencing and direct RNA sequencing. The basic concept involves using an applied voltage to drive single-stranded DNA molecules through a narrow nanopore, which separates chambers of electrolyte solution. This voltage also drives a flow of electrolyte ions through the pore, measured as an electric current. When molecules pass through the nanopore they modify the flow of ions, and structural information can be extracted by analysis of the duration and magnitude of the resulting current reductions. The proposed nanochannel system solves two issues relating to DNA sequencing with solid-state nanopores: 1) feeding long strands of DNA to the sensing element in a single-stranded conformation; 2) reducing the variability found in DNA translocation signals by decreasing the conformation variance of DNA within the nanopore interior. Specifically, we seek to make solid-state ionic-current based nanopore sequencing possible by combining three important components: a nanochannel with sufficiently tight dimensions to allow long strands of DNA to enter the sensing nanopore in an ideal conformation, ultra-thin nanopores to increase signal-to-noise and reduce the number of bases within the pore interior, and optimally fast measurement of translocation through these pores with low-noise, high-bandwidth electronics. Our approach aims to eliminate the need for any enzymes and enables DNA molecules to be geometrically constrained and controlled as they are guided to the nanopores.

项目摘要 为了改善DNA测序并开发RNA测序的实用方法，该NHGRI 第一阶段项目的重点是使用固态纳米孔传感器，宽30nm 嵌入硝酸硅的通道，电子设备在10 MHz带宽下运行DNA 测序和直接RNA测序。基本概念涉及使用施加的电压 驱动单链DNA分子通过狭窄的纳米孔，该纳米孔分开 电解质溶液。该电压还驱动电解质离子通过孔的流动，测量 作为电流。当分子穿过纳米孔时，它们会改变离子的流动， 可以通过分析持续时间和幅度来提取结构信息 导致当前减少。拟议的纳米通道系统解决了与DNA有关的两个问题 用固态纳米孔测序：1）将DNA的长链馈入感应元件中 单链构象； 2）通过 减少纳米孔内部DNA的构象方差。具体来说，我们寻求 通过组合三个 重要组成部分：具有足够紧密尺寸的纳米通道以允许长长 DNA以理想的构型，超薄的纳米孔进入传感纳米孔以增加 信号到噪声并减少孔内内部内的碱数量，并最佳快速 通过这些孔通过这些毛孔进行易位测量，具有低噪声，高带宽电子设备。 我们的方法旨在消除对任何酶的需求，并使DNA分子成为 当它们被引导到纳米孔时，几何受限和控制。