99.99% Accuracy Direct DNA Sequencing via the Protein Nanopore Method

项目摘要 我们的目标是开发一种基于阻断DNA中的离子电流的DNA测序装置。 跨膜蛋白孔<$-溶血素（<$HL）。与其他提出的基于纳米孔的 方法中，蛋白质孔电流阻断（PPCB）方法可以说是最简单的， 概念上和技术上，但最近已经通过了更复杂的支持 方法.提出者的最新电子读出和双层类脂膜进展 已经极大地减轻了先前的信噪比和鲁棒性问题。新的实验数据 和一个精确的DNA运动随机模型（SMDM）在<$HL内现在指示阈值 通过PPCB方法测序的可行性。 对已知输入DNA序列的计算的SMDM响应的检查表明， 通过PPCB方法测序的主要问题是碱基顺序的随机方差 通过孔隙，导致三个可量化的误差来源。该计划将使 对本装置的具体结构和测量参数的修改， 减少每种类型的错误并产生最小的总体测序错误。最终 将通过对高达30 M碱基的天然DNA进行测序来评估仪器。

Project Summary Our aim is to develop a DNA sequencing device based upon the blockade of ionic current in the transmembrane protein pore ¿-hemolysin (¿HL). 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 ¿HL 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 up to 30 Mbases of natural DNA.