NEW METHODS FOR SYNTHESIS OF FLUORESCENT DNA

目标是开发用于直接化学物质的试剂和方法 用作DNA杂交探针的荧光DNA的合成 并进行了引物定向DNA测序。最近，胡德和同事们 开发了一种引物定向DNA测序技术。DNA探针也有 已被发现对检测细菌和病毒以及 基因异常。目前有几家公司正在开发直接 在这种应用中使用的荧光探针的最终目标是 提供快速、灵敏和可靠的临床检测。目前，大多数 制备荧光DNA探针的合成方法依赖于二次 酶和化学修饰。这些发展将会加快 通过介绍直接合成所需产品的方法。 在第一阶段，我们建议合成新的荧光标记核苷酸 适用于自动DNA合成器的亚磷酰胺。这些 化合物将用于合成荧光寡核苷酸。这个 这类化合物的普遍可获得性应会大大刺激 荧光探针在非同位素诊断和治疗中的应用进展 测序试剂。

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.