Electrokinetic Method For Single Molecule DNA Sequencing

单分子 DNA 测序的动电法

• 批准号: 6882886
• 负责人: JOHN G. K. WILLIAMS
• 金额: $ 10万
• 依托单位: LI-COR BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882886
• 关键词: biotechnology; colloids; electrodes; fluorescent dye /probe; high throughput technology; method development; nanotechnology; nucleic acid sequence; oligonucleotides

DESCRIPTION (provided by applicant): We propose a new approach to single-molecule DMA sequencing that can be done in a simple microtiter plate fitted with two electrodes. Over two-hundred different single DMA molecules would be sequenced simultaneously in a single well (maximum 10 nt/s per molecule) with read lengths 20 kb or more. Sequencing cost is potentially lower with this method compared to other long read approaches because of a high degree of multiplexing and because a simple microtiter plate substitutes for expensive micro or nanofabricated devices. While novel in concept, the proposed "electrokinetic" sequencing method would utilize modified polymerases and phosphate-labeled nucleotides from ongoing research on an alternative "charge-switch" method of single molecule sequencing. Ultrabright fluorescent nanoparticles and a prototype two-electrode sequencing chamber will be developed in Phase I. Particle transport kinetics will be characterized using the prototype chamber.

描述（由申请人提供）： 我们提出了一种新的单分子 DMA 测序方法，可以在配有两个电极的简单微量滴定板上完成。超过 200 个不同的单个 DMA 分子将在一个孔中同时进行测序（每个分子最大 10 nt/s），读取长度为 20 kb 或更长。与其他长读长方法相比，这种方法的测序成本可能更低，因为高度多重化，并且简单的微量滴定板取代了昂贵的微米或纳米制造设备。虽然概念新颖，但所提出的“电动”测序方法将利用来自正在进行的单分子测序替代“电荷开关”方法研究中的修饰聚合酶和磷酸盐标记核苷酸。 第一阶段将开发超亮荧光纳米颗粒和原型两电极测序室。将使用原型室来表征粒子传输动力学。