DNA Sequencing by Charge Reduction ESI-MS

通过电荷还原 ESI-MS 进行 DNA 测序

• 批准号: 6436937
• 负责人: LLOYD M SMITH
• 金额: $ 32.74万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6436937
• 关键词: DNA; analytical method; biomedical equipment; biomedical equipment development; electrophoresis; electrospray ionization mass spectrometry; gas analyzer; ions; nucleic acid sequence; particle beam; technology /technique development

DESCRIPTION (provided by applicant): The goal of this proposal is the continued development of DNA sequencing by charge reduction electrospray ionization. The research presented here focuses on the development of new techniques for the generation ionization and sampling of gas-phase biomolecular ions. The combination of these techniques holds the potential to produce a l0,000 fold increase in sensitivity over conventional electrospray. Single droplets will be generated by a piezoelectric dispenser and held in an electrodynamic trap where they will be desolvated by a flow of bath gas to produce analyte ions. The analyte ions will leave the trap along a defined trajectory to enter a charge reduction chamber incorporating aerodynamic lensing. The aerodynamic lens system will replace the standard nozzle-skimmer and collisional cooling systems commonly employed in ESI-TOF. The fact that the ions will travel along a single defined trajectory, rather than along a multitude of trajectories as in conventional electrospray will permit time collection efficiency of the ions into time mass spectrometer to be greatly increased. In principle, the efficiency with which ions are introduced mass spectrometer may approach unity representing an approximately 10,000 fold increase over conventional electrospray. Additionally techniques to increase the ionization efficiency of DNA through gas phase reactions and charge tagging will be researched. The focus of this proposal will be the implementation of the basic concepts outlined above. To this end we will utilize aim existing ESI-TOF mass spectrometer (Perseptive Biosystems Mariner an orthogonal-TOF design) in our laboratory. The use of this existing instrument will eliminate the need to design and build an entirely new mass spectrometer from scratch while permitting dramatic improvements in nmass spectrometric performance.

描述（由申请人提供）：该提案的目的是继续 通过电荷还原电喷雾电离开发DNA测序。这 这里介绍的研究重点是开发 气相生物分子离子的产生电离和采样。这 这些技术的组合具有产生L0,000倍的潜力 对常规电喷雾的灵敏度提高。单滴是 由压电分配器产生，并保持在电动陷阱中 它们将被沐浴气体的流动降低，以产生分析物离子。这 分析物离子将沿定义的轨迹离开陷阱以输入电荷 还原室，融合了空气动力学镜头。空气动力学镜头 系统将替换标准的喷嘴蒸馏器和碰撞冷却系统 通常在ESI-TOF中使用。离子将沿着一个单一行进的事实 定义的轨迹，而不是沿着多种轨迹 常规电喷雾将允许离子的时间收集效率 到时间质谱仪将大大增加。原则上， 引入离子质谱仪的效率可能接近统一 代表比常规的大约10,000倍 电喷雾。另外提高电离效率的技术 将研究通过气相反应和电荷标记的DNA。这 该建议的重点将是实施基本概念 上面概述了。为此，我们将利用AIM现有的ESI-TOF质量 光谱仪（感染性生物系统水手是正交TOF设计） 实验室。这种现有工具的使用将消除 从头开始设计和建造一个全新的质谱仪 允许NMASS光谱性能的显着改善。