99.99% Accuracy Direct DNA Sequencing via the Protein Nanopore Method

99.99%%20准确度%20直接%20DNA%20测序%20via%20the%20蛋白质%20纳米孔%20方法

• 批准号: 7714702
• 负责人: Geoffrey Barrall
• 金额: $ 96.15万
• 依托单位: ELECTRONIC BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7714702
• 关键词: Algorithms; Amino Acid Sequence; Award; Back; Base Sequence; Basic Science; Biological Process; Code; Complex; Custom; DNA; DNA Modification Process; DNA Sequence; Data; Data Analyses; Devices; Diagnostic; Diffusion; Electrolytes; Electronics; Elements; Error Sources; Forensic Medicine; Genome; Grant; Hemolysin; Integral Membrane Protein; Link; Lipid Bilayers; Location; Measurement; Measures; Medical; Methods; Modeling; Modification; Motion; Mutation; Noise; Operative Surgical Procedures; Peptide Sequence Determination; Performance; Polymers; Pore Proteins; Process; Proteins; Protocols documentation; Relative (related person); Research; Sampling; Scheme; Signal Transduction; Single-Stranded DNA; Site; Site-Directed Mutagenesis; Structure; System; Technology; Testing; Variant; base; biological research; cost; insight; instrumentation; mathematical model; nanopore; programs; public health relevance; reconstruction; research study; response; simulation; statistics; voltage

Our aim is to develop a DNA sequencing device based upon the blockade of ionic current in the transmembrane protein pore alpha-hemolysin (aHL). Compared to other proposed nanopore-based approaches, the protein pore current blockade (PPCB) method is arguably the simplest, both conceptually and technologically, but recently has been passed over in favor of more complex methods. Recent electronic readout and bilayer lipid membrane advances by the proposers have greatly alleviated prior signal-to-noise ratio and robustness issues. New experimental data and an accurate Stochastic Model for DNA Motion (SMDM) within aHL now indicate threshold feasibility for sequencing by the PPCB method. Inspection of calculated SMDM responses to known input DNA sequences shows that the principal issue for sequencing via the PPCB method is random variance in the order the bases pass through the pore, leading to three quantifiable sources of error. This program will make specific structural and measurement parameter modifications to the present apparatus to reduce each type of error and to produce a minimal overall sequencing error. The final apparatus will be evaluated by sequencing kilobase strands of natural DNA

我们的目标是开发一种基于离子电流阻断的DNA测序装置 在跨膜蛋白孔中的α-溶血素（阿勒）。与其他建议相比 基于纳米孔的方法，蛋白质孔电流阻断（PPCB）方法是 可以说是最简单的，无论是概念上还是技术上，但最近已经 而不是更复杂的方法。最近的电子读出和双层 脂膜的进步，由提议者大大减轻了先前的信号噪声 比率和鲁棒性问题。新的实验数据和精确的随机模型 阿勒内的DNA运动（SMDM）现在表明通过以下方法进行测序的阈值可行性： PPCB方法。对已知输入DNA的计算SMDM响应的检查 序列表明，通过PPCB方法测序的主要问题是随机的 碱通过孔的顺序的变化，导致三个可量化的来源 的错误。本程序将制定具体的结构和测量参数 为了减少每种类型的误差并产生 最小的整体测序误差。将通过测序对最终装置进行评价 天然DNA的内切酶链