Multiplex PCR-based Alu insertion polymorphisms genotyping for identifying individuals of Japanese ethnicity
Multiplex PCR-based Alu insertion polymorphisms genotyping for identifying individuals of Japanese ethnicity
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基于多重 PCR 的 Alu 插入多态性基因分型用于识别日本种族个体
DOI：
10.1016/j.ygeno.2012.01.004
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发表时间：
2012
期刊：
影响因子：
4.4
通讯作者：
K. Shimizu
中科院分区：
文献类型：
--
作者：
M. Asari;T. Omura;K. Oka;C. Maseda;Y. Tasaki;H. Shiono;K. Matsubara;M. Matsuda;K. Shimizu
Alu插入片段的识别是地理谱系分析的有用工具，通常通过Alu元素扩增和琼脂糖凝胶电泳法进行。在这里，我们开发了一种新的基于荧光的法医鉴定中多重Alu基因分型的方法。我们检测了70名日本人的等位基因频率，并选择了30个多态的Alu插入。为每个Alu基因座设计3个引物，利用扩增片段大小3~6bp的差异来区分等位基因。此外，我们还将30个基因座的扩增引物分成三组，每组引物扩增出10个片段，片段大小在50~137bp之间。根据群体数据，错误分配匹配的概率为3.7x10DNA 13。三次独立扩增和随后的毛细管电泳能对少量−进行灵敏的基因分型，表明该方法适用于识别日裔个体。
Discrimination of Alu insertions is a useful tool for geographic ancestry analysis, and is usually performed by Alu element amplification and agarose gel electrophoresis. Here, we have developed a new fluorescence-based method for multiple Alu genotyping in forensic identification. Allele frequencies were determined in 70 Japanese individuals, and we selected 30 polymorphic Alu insertions. Three primers were designed for each Alu locus to discriminate alleles using the 3–6bp differences in amplicon sizes. Furthermore, we classified the amplification primers for the 30 loci into three different sets, and PCR using each set of primers provided 10 loci fragments ranging from 50 to 137bp. Based on population data, the probability of incorrectly assigning a match was 3.7×10−13. Three independent amplifications and subsequent capillary electrophoresis enabled the sensitive genotyping of small amounts of DNA, indicating that this method is suitable for identifying individuals of Japanese ethnicity.