ELECTROPHORE LABELS FOR DNA PROBES

这个项目的目的是开发新的分析方法 利用电泳仪来加快基因图谱的绘制。 电泳石是一种在气体中检测到的挥发性化合物 通过电子俘获技术进行相变。我们已经设计了一种方法来使用 作为DNA探针的标记的电泳膜。最重要的属性是 出于这一目的，电泳仪具有广泛的应用潜力 多样性。这指的是利用许多 在给定的化验中，同时作为标记的电泳膜。例如, 许多不同的DNA探针中的每一个都可以用唯一的 电泳液，然后组合成多探针鸡尾酒。 我们建议开发基于以下因素的遗传作图方法 用凝胶标记的DNA探针进行杂交分析。化验结果 将以点印迹的形式执行，其中每个印迹都与 含有许多这样的探针的溶液。DNA的许多靶序列 因此，可以同时测试斑点印迹。 更详细地说，我们在这个创业项目中提议建立一个 电泳法标记的DNA探针的重复性为12。这些探头 将被组合并用于测试Bluescrip、lambda、 PhiX174和腺病毒DNA。探测将由电子完成 利用粒子束捕获负离子质谱仪(ECNI-MS 接口的吞吐量为15秒/样本。一种气相色谱 (GC)接口也将被研究。 还提出了未来工作的计划，超出了本章的范围 启动项目，其中的多样性将被提升到 范围在100到400之间，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种 每小时24,000次杂交分析的吞吐量 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.