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

• 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
• 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' -deoxyguanosine 5' -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

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，使用一对用包含氢键供体和氢键受体的简单试剂功能化的隧道电极，可以为所有四种核苷（和5-甲基脱氧胞苷）产生不同的隧道信号。 该提案的目标是将测量扩展到水性电解质中的核苷酸，然后扩展到小寡聚物。 我们将量化单分子读数的分数，并确定控制这一分数的因素，目的是消除一次来自间隙中多个核苷酸的信号。 我们将探索控制所有四个碱基（和 5-甲基 C）电流信号分布宽度的因素，目的是提高单次读取的辨别力。 我们将测量成功读取的比例，并表征复合物（产生信号）在隧道间隙中形成所需的时间。 从这些测量中，我们将确定提高读出效率所需的改进，并制定配备隧道电极的纳米孔测序系统的设计标准。 在这项研究过程中开发的试剂将可供其他研究小组使用，这些研究小组正在开发使用电子隧道作为读数的纳米孔测序仪。 公共健康相关性：至少有七个 NIH 支持的团体正在探索测序方法，建议使用电子隧道作为纳米孔测序仪的读数，这种方法可能会大大降低测序成本。 我们已经证明，所有四种核苷和 5-甲基胞苷都可以通过功能化电极读取，我们将开发适合在水性电解质中进行 DNA 测序的试剂，并使这些试剂广泛使用。