The selection of new microsatellite loci suitable for fortune forensic exam ination and the establishment of its wide use

适合命理检验的新微卫星位点的筛选及其广泛应用的建立

基本信息

批准号：
10557046
负责人：
KATSUMATA Yoshinao
金额：
$ 7.94万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557046/
关键词：
microsatellite STR capillary electrophoresis fluorescent labeled primer allele frequency heterozygosity

项目摘要

Within the term of this research project (2 years), we analyzed 168 STR loci spacing at 25 cM on average in human genome using a STR primer set for linkage analysis, a MapPair 8A (Research Genetics Inc.). This primer set includes 168 primer pairs in which one of each pair is labeled with either of the tricolor fluorescent dyes (FAM, HEX, TET). DNA samples were amplified and the PCR products were electrophoresed according to the Version 8A multiplex table with the Genetic Analyzer 310, and each sample was typed from sizing results. We have finished to analyze all 168 loci with 32 Japanese DNA samples. Out of 20 STR loci showing heterozygosities of higher than 0.8 twelve STR loci were selected because their sequence structures were simple based on the databases, GDB etc. The sequences of those loci were confirmed accurately in this study with minor corrections.We tried to construct 2 multiplex systems with each 6 STR loci using the tricolor fluorescent dyes. We newly designed each primer set at each locus for the new systems. One system was completed and another has almost been completed. Moreover, we have also devised a mew multiplex system using 3 STR loci (wglc5, wgld1, pq2h2) with comparatively high heterozygosity and without irregular allele selected from approximately 20 microsatellite loci which Prof. Armour in Nottingham University had isolated. The results including the usefulness in the forensic field were presented at the 8th annual meeting of the Japanese Society for DNA polymorphism Research.In this way, the new microsatellite typing systems using 15 loci appropriate to forensic examination in Japanese has almost been completed.

在该研究项目期间（2 年），我们使用用于连锁分析的 STR 引物组 MapPair 8A（Research Genetics Inc.）分析了人类基因组中平均间距为 25 cM 的 168 个 STR 位点。该引物组包括 168 个引物对，其中每对中的一个用三色荧光染料（FAM、HEX、TET）之一进行标记。使用遗传分析仪 310 根据版本 8A 多重表对 DNA 样本进行扩增，并对 PCR 产物进行电泳，并根据大小结果对每个样本进行分类。我们已经完成了 32 个日本 DNA 样本的全部 168 个基因座的分析。在 20 个杂合度高于 0.8 的 STR 位点中，选择了 12 个 STR 位点，因为基于数据库、GDB 等，它们的序列结构很简单。这些位点的序列在本研究中经过较小的修正得到了准确的确认。我们尝试构建 2使用三色荧光染料，每个 6 个 STR 位点的多重系统。我们为新系统的每个基因座新设计了每个引物组。一个系统已经完成，另一个系统也即将完成。此外，我们还设计了一个mew多重系统，使用3个STR基因座（wglc5，wgld1，pq2h2），杂合性相对较高，并且没有不规则等位基因，选自诺丁汉大学Armour教授分离的约20个微卫星基因座。包括在法医领域的实用性在内的研究结果已在日本DNA多态性研究学会第八届年会上公布。这样，使用适合日语法医检查的15个基因座的新微卫星分型系统已基本完成。