DNA SEQUENCING BY CHARGE REDUCTION ESI-MS

通过电荷还原 ESI-MS 进行 DNA 测序

• 批准号: 2889702
• 负责人: LLOYD M SMITH
• 金额: $ 12.59万
• 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 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2889702
• 关键词: analytical method; biomedical equipment development; electrospray ionization mass spectrometry; mass spectrometry; nucleic acid sequence

DESCRIPTION: (Applicant's Abstract) In this project we propose to develop a novel mass spectrometric method, Charge Reduction (CR) Electrospray Ionization (ESI) Mass Spectrometry (MS), for the analysis of enzymatic DNA sequencing reactions. A nanospray Charge Reduction source will be constructed and interfaced with a commercial Electrospray Time-of-Flight (ESI-TOF) instrument. This source will utilize the emitting radioisotope 210Po to ionize the source bath gas, producing ions which in turn neutralize the charges present on the ions generated in the ESI process. This charge neutralization process will greatly reduce the complexity of the mass spectra obtained in ESI-MS from large biopolymers such as DNA, thereby enabling the mass spectrometric analysis of biopolymer mixtures which are otherwise too complex to be useful. It is anticipated that the ability to reduce the number of charge states produced in the electrospray method, coupled with the intrinsic speed and accuracy of mass spectrometric analysis, will provide a viable route to the rapid mass spectrometric analysis of Sanger sequencing reactions in seconds, a dramatic improvement over the capabilities of electrophoretic methods.

描述：（申请人的摘要）在这个项目中，我们建议开发一个 新型质谱法，电荷还原（CR）电喷雾 电离（ESI）质谱法（MS），用于分析酶促DNA 测序反应。 纳米喷的减少源将是 与商业电喷雾飞机构建并连接 （ESI-TOF）仪器。 该来源将利用发射的放射性同位素 210po使源浴气电离，产生离子又中和 ESI过程中产生的离子上存在的电荷。 这是费用 中和过程将大大降低质量的复杂性 从大型生物聚合物（例如DNA）中获得的光谱，从而 实现生物聚合物混合物的质谱分析 否则太复杂了，无法有用。 可以预期 减少电喷雾方法中产生的电荷状态的数量， 再加上质谱的内在速度和准确性 分析将为快速质谱法提供可行的途径 分析桑格测序反应以秒为单位的反应，这是一个显着的改进 电泳方法的能力。