Tunnel Junction for reading all four bases with high discrimination

隧道连接，用于以高辨别力读取所有四个碱基

基本信息

批准号：
8134442
负责人：
STUART LINDSAY
金额：
$ 28.92万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8134442
关键词：
2&apos -deoxyguanosine 5&apos -phosphate Binding Buffers Characteristics Chemicals Chemistry Complex Cytidine DNA DNA Sequence Data Deoxycytidine Detection Development Discrimination Electrodes Electrolytes Electrons Elements Environment Generic Drugs Goals Head Hemolysin Hydrogen Bonding Ion Channel Ions Learning Measurement Measures Methods Nature Noise Nucleosides Nucleotides Physics Polymers Proteins Reader Reading Reagent Relative (related person)Research Sampling Signal Transduction Solutions Solvents Speed Structure Support Groups System Theoretical model Time United States National Institutes of Health Width Work Writing aqueous base cost design electric field improved mutant nanopore operation public health relevance reagent testing research study response simulation single molecule

2&apos -deoxyguanosine 5&apos -phosphate Binding Buffers Characteristics Chemicals Chemistry Complex Cytidine DNA DNA Sequence Data Deoxycytidine Detection Development Discrimination Electrodes Electrolytes Electrons Elements Environment Generic Drugs Goals Head Hemolysin Hydrogen Bonding Ion Channel Ions Learning Measurement Measures Methods Nature Noise Nucleosides Nucleotides Physics Polymers Proteins Reader Reading Reagent Relative (related person)Research Sampling Signal Transduction Solutions Solvents Speed Structure Support Groups System Theoretical model Time United States National Institutes of Health Width Work Writing aqueous base cost design electric field improved mutant nanopore operation public health relevance reagent testing research study response simulation single molecule

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项目摘要

DESCRIPTION (provided by applicant): We have discovered that distinct tunneling signals can be generated for all four nucleosides (and 5-methyldeoxycytidine) using one pair of tunneling electrodes functionalized with a simple reagent containing a hydrogen-bond donor and a hydrogen bond acceptor. The goals of this proposal are to extend the measurements to nucleotides in aqueous electrolyte, and then to small oligomers. We will quantify the fraction of single-molecule reads and determine the factors that control this fraction with the goal of eliminating signals that come from more than one nucleotide in the gap at a time. We will explore the factors that control the width of the distribution of current signals for all four bases (and 5-methyl C) with the goal of improving the discrimination of a single read. We will measure the fraction of successful reads and characterize the time required for the complex (that gives rise to the signal) to form in the tunnel gap. From these measurements, we will identify improvements needed to increase the readout efficiency and also develop criteria for design of a nanopore sequencing system equipped with tunneling electrodes. The reagents developed during the course of this research will be made available to other research groups developing nanopore sequencers that use electron tunneling as the readout. PUBLIC HEALTH RELEVANCE: At least seven NIH-supported groups are exploring sequencing methods that propose to use electron tunneling as the readout for a nanopore sequencer, an approach that might greatly reduce the cost of sequencing. We have shown that all four nucleosides and 5-methyl cytidine can be read by functionalized electrodes and we will develop reagents suitable for DNA sequencing in aqueous electrolyte and make these widely available.

描述（由申请人提供）：我们发现，可以使用一对用一对含有氢键供体和氢键受体的简单试剂功能化功能化的隧穿电极来为所有四种核苷（和5-甲基氧基辛丁丁）生成不同的隧穿信号。该建议的目标是将测量值扩展到水溶液中的核苷酸，然后将其扩展到小寡聚物中。我们将量化单分子读取的分数，并确定控制这一分数的因素，以消除一次间隙中多个核苷酸的信号。我们将探讨控制所有四个基碱基（和5-甲基C）当前信号分布宽度的因素，以改善单个读取的歧视。我们将测量成功读取的分数，并表征复合物（引起信号）在隧道间隙中形成所需的时间。从这些测量值中，我们将确定提高读数效率所需的改进，并制定配备有隧道电极的纳米孔测序系统设计标准。在本研究过程中开发的试剂将提供给开发使用电子隧道作为读数的纳米孔测序仪的其他研究组。公共卫生相关性：至少有七个由NIH支持的组探索了测序方法，这些方法建议将电子隧道用作纳米孔序列的读数，这种方法可能会大大降低测序的成本。我们已经表明，所有四种核苷和5-甲基胞丁都可以通过功能化的电极读取，我们将开发适合于水解中DNA测序的试剂，并使其广泛可用。