DNA SEQUENCING BY LASER-BASED, MASS SPECTRAL METHODS

该提案概述了一个专注于使用最先进技术的计划 解决重要生物学问题的实验方法。 的 RCDA 提案的近期目标是开发新的高速 DNA测序仪。 该仪器的实现将使DNA 测序速度接近每百万个碱基 day - 速率比当前速率快三个数量级 技术。 为了实现这一目标，当前的 DNA 测序策略 将与三项新技术相结合，生产出一种低成本、容易 自动测序方法。 激光汽化用于转移标记 DNA 链进入气相。 一旦进入气相共振—— 增强的多光子电离用于精确放置一个单位 每条链上带正电荷，然后进行飞行时间质量分析 执行。 所提议技术的自动化将导致 极其灵敏且具有成本效益的 DNA 测序方法需要 很少的体力劳动。

This proposal outlines a program focused on using state-of-the-art experimental methods to address important biological problems. The immediate goal of this RCDA proposal is the development of a new high speed DNA sequencer. The realization of this instrument will allow DNA sequencing to be performed at a rate approaching one million bases per day - a rate three orders of magnitude faster than currently available techniques. To achieve this objective current DNA sequencing strategies will be combined with three new technologies to produce a low-cost, easily automated sequencing method. Laser vaporization is used to transfer tagged strands of DNA into the gas phase. Once in the gas phase resonance- enhanced, multiphoton ionization is used to place precisely one unit of positive charge on each strand and then time-of-flight mass analysis performed. The automation of the technology proposed would result in an extremely sensitive and cost effective DNA sequencing method requiring little manual labor.