ELECTROPHORE LABELS FOR DNA PROBES

本项目的目的是开发新的分析方法 让生物学家参与加速基因图谱的绘制。 电泳体是在气体中检测到的挥发性化合物 通过电子捕获技术相。 我们发明了一种方法 作为DNA探针的标记物。 最重要的财产 这一目的的重要性在于它们具有广泛的 多样性 这是指利用许多机会 在给定的测定中同时作为标记。 比如说， 许多不同的DNA探针中的每一种都可以用独特的 将该混合物与电泳团混合，然后组合用作多探针混合物。 我们建议发展遗传作图的方法学， 用荧光标记的DNA探针进行杂交测定。 的测定 将以斑点印迹进行，其中每个印迹与 含有许多这样的探针的溶液。 DNA中的许多靶序列 从而可以同时测试斑点印迹。 更详细地说，我们建议在这个启动项目中建立一个 12的多重性，用于标记的DNA探针。 这些探针 将被组合并用于测试Bluescript，lambda， phiX 174和腺病毒DNA。 探测将由电子 使用粒子束的捕获负离子质谱（ECNI-MS） 以15秒/样品的吞吐量连接。 气相色谱 (GC)接口也将进行研究。 此外，还介绍了本报告范围以外的未来工作计划。 启动项目，其中多重性将被提升到 范围为100至400，同时保持相同的分析速度15 秒/点印迹。 这将为检测步骤提供一个 每小时24，000次杂交测定的通量， 100.

The purpose of this project is to develop new analytical methodology involving electrophores to speed up genetic mapping. An electrophore is a volatile compound which is detected in the gas phase by an electron capture technique. We have devised a means to use electrophores as labels for DNA probes. The most important property of electrophores for this purpose is their potential for extensive multiplicity. This refers to the opportunity to utilize many electrophores simultaneously as labels in a given assay. For example, each of many different DNA probes could be labeled with a unique electrophore, and then combined for use as a multi-probe cocktail. We propose to develop methodology for genetic mapping based on hybridization assays with electrophore-labeled DNA probes. The assays will be performed as dot blots in which each blot is hybridized with a solution containing many such probes. Many target sequences of DNA in the dot blot can thereby be tested simultaneously. In more detail, we propose in this startup project to establish a multiplicity of 12 for electrophore-labeled DNA probes. These probes will be combined and used to test do blots of Bluescript, lambda, phiX174 and adenovirus DNA. Detection will be accomplished by electron capture negative ion mass spectrometry (ECNI-MS) using a particle beam interface at a throughput of 15 seconds/sample. A gas chromatography (GC) interface will also be studied. Also presented are plans for future work, beyond the scope of this startup project, in which the multiplicity would be raised into the range of 100 to 400 while maintaining the same analysis speed of 15 seconds/dot blot. This would provide, for the detection step, a throughput of 24,000 hybridization assays per hour for a multiplicity of 100.