SEQUENCING OF DNA BY LASER IONIZATION

通过激光电离进行 DNA 测序

• 批准号: 2674189
• 负责人: CHRISTOPHER H BECKER
• 金额: $ 27.8万
• 依托单位: GENETRACE SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2674189
• 关键词: DNA; biomedical equipment development; biotechnology; ionization; laser spectrometry; mass spectrometry; nucleic acid sequence

DESCRIPTION (Adapted from the Investigator's Abstract): The principal goal of this research is to further develop laser desorption/ionization time-of-flight mass spectrometry into a practical technique for high-speed DNA sequencing and sizing. Specific technical aims include unit base resolution in mixtures of DNA oligomers containing up to 300 nucleotides with sensitivity levels of less than or equal to one femtomole component. A reduction in molecular fragmentation and adduct formation in addition to multiply charged ions and dimer ions is also required. Central to the success of the desorption/ionization of fragile DNA oligomers is the nature of the matrix and its preparation with the oligonucleotides which must be optimized in order to achieve the technical goals. Proposed work will emphasize novel preparation techniques using ultraviolet-sensitive matrices such as 3-hydroxypicolinic acid as well as cryogenic frozen solutions for new matrices. Several improvements in instrumentation will be studied to further improve performance with existing and new sample preparation methods. While most of the research will be directed toward improving the performance of the basic mass spectrometry for oligonucleotides, a small fraction of the effort will be devoted to demonstrating genomics applications, including those where the high speed of the mass spectrometry can be used to advantage even with the current state-of-the-art technology.

描述（改编自研究者摘要）：主要目标 进一步发展激光解吸/电离 将飞行时间质谱技术应用于高速 DNA测序和大小。 具体技术目标包括单位基数 含有多达300个核苷酸的DNA寡聚体混合物的分辨率 灵敏度水平小于或等于一个飞摩尔分量。 一 减少分子断裂和加合物的形成， 还需要多电荷离子和二聚体离子。 脆性DNA寡聚体解吸/电离成功的关键 基质的性质及其与寡核苷酸的制备 其必须被优化以实现技术目标。 提出 工作将强调新的制备技术， 基质如3-羟基吡啶甲酸以及低温冷冻 新矩阵的解决方案。 将对仪器进行几项改进， 研究以进一步提高现有和新样品的性能 制备方法 虽然大多数研究将针对 改进基本质谱的性能， 寡核苷酸，一小部分的努力将致力于 展示基因组学的应用，包括那些 即使在电流 最先进的技术

项目成果

Cryogenic frozen solution matrixes for analysis of DNA by time-of-flight mass spectrometry.

用于通过飞行时间质谱分析 DNA 的低温冷冻溶液基质。

• DOI: 10.1021/ac9703764
• 发表时间: 1997
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Hunter,JM;Lin,H;Becker,CH
• 通讯作者: Becker,CH

High-throughput DNA analysis by time-of-flight mass spectrometry.

通过飞行时间质谱法进行高通量 DNA 分析。

• DOI: 10.1038/nm0397-360
• 发表时间: 1997
• 期刊: Nature medicine
• 影响因子: 82.9
• 作者: Monforte,JA;Becker,CH
• 通讯作者: Becker,CH