Cenanović Merisa, Pojskić Naris, Kovacević Lejla, Dzehverović Mirela, Cakar Jasmina, Musemić Dzenisa, Marjanović Damir
Institute for Genetic Engineering and Biotechnology, Sarajevo University, Sarajevo, Bosnia and Herzegovina.
Coll Antropol. 2010 Jun;34(2):545-50.
In our previous population study, we have used twelve Y-chromosomal short tandem repeats loci incorporated in the PowerPlex Y System to determine Y-STR diversity in B&H human population. With intent to obtain additional verification of the previously obtained results as well as to establish specific reference for a local B&H population, we have decided to test DNA samples collected from 100 unrelated healthy male Canton Sarajevo residents (from Sarajevo region) for the same twelve Y-linked short tandem repeats loci. Qiagen DNeasy Tissue Kit (Qiagen, GmbH, Hilden, Germany) was used for DNA extraction from buccal swabs and PowerPlex Y System (Promega Corp., Madison, WI) has been used to simultaneously amplify Y-STR loci by PCR. PowerPlex Y System includes 12 STR loci: DYS19, DYS385a, DYS385b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439. The total PCR reaction volume was 5 microL. PCR amplifications were carried out in PE GeneAmp PCR System Thermal Cycler (ABI). Electrophoresis of the amplification products was preformed on an ABI PRISM 310 genetic analyzer (ABI, Foster City, CA) according to the manufacturer's recommendations. The raw data were compiled and analyzed using the accessory software: ABI PRISM Data Collection Software and Genemapper version 3.2. In addition, we have compared the obtained "Sarajevo" dataset with the data previously generated for the entire Bosnian and Herzegovinian population, as well as with the available data on geographically close (neighboring) European populations. The results of this study will be used as guidelines in additional improving of research into genetic relationship among recent local B&H populations, both isolated and open, which is a long-term project in our country.
在我们之前的人群研究中,我们使用了PowerPlex Y系统中包含的12个Y染色体短串联重复序列位点来确定波黑人群中的Y-STR多样性。为了进一步验证之前获得的结果,并为当地的波黑人群建立特定的参考标准,我们决定对从100名来自萨拉热窝地区、互不相关的健康男性萨拉热窝市民采集的DNA样本进行相同的12个Y连锁短串联重复序列位点检测。使用Qiagen DNeasy Tissue试剂盒(Qiagen公司,德国希尔德)从口腔拭子中提取DNA,并使用PowerPlex Y系统(Promega公司,美国威斯康星州麦迪逊)通过PCR同时扩增Y-STR位点。PowerPlex Y系统包括12个STR位点:DYS19、DYS385a、DYS385b、DYS389I、DYS389II、DYS390、DYS391、DYS392、DYS393、DYS437、DYS438和DYS439。PCR反应总体积为5微升。PCR扩增在PE GeneAmp PCR系统热循环仪(ABI)中进行。扩增产物的电泳在ABI PRISM 310基因分析仪(ABI公司,美国加利福尼亚州福斯特城)上按照制造商的建议进行。原始数据使用配套软件ABI PRISM数据收集软件和Genemapper 3.2版进行整理和分析。此外,我们将获得的“萨拉热窝”数据集与之前为整个波斯尼亚和黑塞哥维那人群生成的数据,以及地理上相近(相邻)的欧洲人群的现有数据进行了比较。本研究的结果将作为进一步改进我国近期孤立和开放的当地波黑人群之间遗传关系研究的指导方针,这是我国的一个长期项目。