PCR-BASED DNA TYPING SYSTEM

我们建议开发一种使用聚合酶链进行DNA分型的方法 反应(PCR)。我们将开发一系列聚合酶链式反应引物， 扩增人类DNA的特定VNTR区。我们将优化条件， 使产生的DNA片段能够在聚丙烯酰胺上可视化 凝胶。通过使用几对聚合酶链式反应引物，一种明确的带型 将为每个人生成。其结果将是一场迅速的、非 可用于法医和亲子鉴定的同位素技术 不同种族人群的等位基因频率。这将允许 排除概率和同一性概率的确定 亲子鉴定、法医鉴定和犯罪研究。 在第一阶段，我们将通过开发 我们的一个高度多态的探针的一对聚合酶链式反应引物 包含vntr。我们将进一步展示我们创造聚合酶链式反应- 通过为不包含该基因的探针开发引物进行测试 Vntr。此外，我们还将优化电泳条件 从我们的基因扩增产物中分离出的片段 模型系统。在第二阶段，我们将完成对 我们的全套亲子鉴定/法医调查建立了一个数据数据库 分析和制定视频成像系统和软件的使用情况 数据收集和存储。

We propose to develop a method of DNA typing using the polymerase chain reaction (PCR). We will develop a collection of PCR primers which will amplify specific VNTR regions of human DNA. we will optimize conditions to enable visualization of the resulting DNA fragments on a polyacrylamide gel. By using several pairs of PCR primers, an unambiguous banding pattern will be generated for each individual. The result will be a rapid, non- isotopic technique that can be used in forensic and paternity analyses and frequency of alleles for different ethnic groups. This will allow determination of probability of exclusion and probability of identity in paternity, forensic and criminal studies. In phase I we will demonstrate the feasibility of this system by developing a pair of PCR primers for one of our highly polymorphic probes that contains the VNTR. We will further demonstrate our ability to create PCR- based tests by developing primers for a probe that does not contain the VNTR. In addition we will optimize conditions for electrophoretic separation of fragments resulting from amplification using primers from our model system. In phase II we will finish development of PCR primers for our entire set of paternity/forensic probes, develop a database for data analysis and institute the use of a video imaging system and software for data collection and storage.