NEW METHODS FOR SYNTHESIS OF FLUORESCENT DNA

荧光 DNA 合成的新方法

• 批准号: 3497835
• 负责人: RICHARD H TULLIS
• 金额: $ 5万
• 依托单位: IMMUNE COMPLEX CORPORATION
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3497835
• 关键词: DNA; communicable disease diagnosis; fluorescence; fluorescent dye /probe; gel electrophoresis; gel filtration chromatography; high performance liquid chromatography; nucleic acid chemical synthesis; nucleic acid sequence; oligonucleotides; phosphamide

The goal is to develop reagents and methods for the direct chemical synthesis of fluorescent DNAs destined for use as DNA hybridization probes and in primer directed DNA sequencing. Recently, Hood and coworkers have developed a primer directed DNA sequencing technique. DNA probes have also been found to be useful for detecting bacteria and viruses as well as genetic abnormalities. Several companies are currently developing direct fluorescent probes for use in such applications with the ultimate goal of producing rapid, sensitive and reliable clinical tests. Currently, most synthetic procedures for making fluorescent DNA probes rely on secondary enzymatic and chemical modification. These developments would be speeded by the introduction of methods for direct synthesis of the desired product. In Phase i we propose to synthesize novel fluorescent labeled nucleotide phosphoramidites suitable for use in automated DNA synthesizers. These compounds will be used to synthesize the fluorescent oligonucleotides. The general availability of such compounds should significantly stimulate progress in the use of fluorescent probes as non-isotopic diagnostic and sequencing reagents.

目标是开发直接化学反应的试剂和方法， 用作DNA杂交探针的荧光DNA的合成 以及引物指导的DNA测序。 最近，胡德和同事们 开发了引物指导的DNA测序技术。 DNA探针也 已发现其可用于检测细菌和病毒， 基因异常 目前有多家公司正在开发直接 用于这些应用的荧光探针的最终目标是 产生快速、灵敏和可靠的临床测试。 目前大多数 用于制备荧光DNA探针的合成方法依赖于二级结构， 酶和化学修饰。 这些发展将会加快 通过引入直接合成所需产物的方法。 在第一阶段，我们提出了合成新的荧光标记的核苷酸， 适用于自动DNA合成仪的亚磷酰胺。 这些 化合物将用于合成荧光寡核苷酸。 的 这些化合物的普遍可用性将显著刺激 荧光探针作为非同位素诊断和 测序试剂。