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细胞内离子流阻断的DNA测序装置 跨膜蛋白孔溶血素(HL)。与其他建议的基于纳米孔的 方法，蛋白质孔电流阻断(PPCB)方法可以说是最简单的，两者 在概念上和技术上，但最近已经被忽略，转而支持更复杂的 方法：研究方法。电子读数和双层类脂膜的最新进展 极大地缓解了先前的信噪比和稳健性问题。新的实验数据 一个精确的DNA运动随机模型(SMDM)在HL内现在表明阈值 多氯联苯测序方法的可行性。 对计算的SMDM对已知输入DNA序列的响应进行检查表明， 通过PPCB法进行测序的主要问题是碱基顺序的随机变化 穿过毛孔，导致三个可量化的误差来源。这一计划将使 对本装置进行特定的结构和测量参数修改以 减少每种类型的误差，并产生最小的总体排序误差。决赛 该仪器将通过对高达30M的天然DNA碱基进行测序进行评估。

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.