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

• 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; microsatellite; STR; capillary electrophoresis; fluorescent labeled primer; allele frequency; heterozygosity

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.

在该研究项目的任期内（2年），我们使用链接分析的STR底漆集，平均分析了人类基因组的168个str座位间距，Mappair 8a（研究Genetics Inc.）。该底漆集包括168对，其中每对都用三色荧光染料（FAM，HEX，TET）标记。放大DNA样品，并根据具有遗传分析仪310的版本的多路复用表对PCR产物进行电泳，并根据尺寸结果键入每个样品。我们已经用32个日本DNA样品分析了所有168个基因座。在20个STR基因座中，显示出高于0.8二十二个基因座的杂合子的序列结构，因为它们的序列结构是根据数据库，GDB等简单的。在本研究中，这些基因座的序列在次要的校正中得到了准确的确认。我们尝试使用Tricolor荧光荧光染料使用6个str座位进行次要校正。我们新设计的每个底漆在每个基因座都为新系统设置。一个系统已经完成，另一个系统几乎已经完成。此外，我们还使用3个str基因座（WGLC5，WGLD1，PQ2H2）设计了一个MEW多路复用系统，其杂合性相对较高，并且没有从诺丁汉大学的Armor教授的大约20个Microsatellite Loci中选择的不规则等位基因。在日本DNA多态性研究协会的第八届年度会议上，介绍了包括法医领域的实用性的结果。这样，使用15个基因座的新的微卫星打字系统几乎完成了。