Genotyping of 38 insertion/deletion polymorphisms for human identification using universal fluorescent PCR

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• 作者：Kumiko Oka ; Masaru Asari ; Tomohiro Omura ; Masatsugu Yoshida ; Chikatoshi Maseda ; Daisuke Yajima ; Kazuo Matsubara ; Hiroshi Shiono ; Mitsuyoshi Matsuda ; Keiko Shimizu
• 关键词：Insertion/deletion polymorphism ; Universal fluorescent PCR ; Human identification ; Forensic genotyping ; Capillary electrophoresis ; Multiplex PCR
• 刊名：Molecular and Cellular Probes
• 出版年：February, 2014
• 年：2014
• 卷：28
• 期：1
• 页码：13-18
• 全文大小：964 K

文摘

Short insertion/deletion (Indel) polymorphisms of approximately 2-6聽bp are useful as biallelic markers for forensic analysis, and the application of Indel genotyping as a supplementary tool would improve human identification accuracy. We examined the allele frequencies of 37 autosomal Indels in the Japanese population and developed a novel dual-color genotyping method for human identification on the basis of universal fluorescent PCR, including the sex-typing amelogenin locus. Target genomic fragment sizes for 38 Indels were 49-143聽bp. We analyzed these Indels in 100 Japanese individuals using the M13(-47) sequence as a universal primer. For dual-color genotyping, we designed a novel universal primer with high amplification efficiency and specificity. Using FAM-labeled M13(-47) and HEX-labeled modified M13(-47) primers, fluorescent signals at all loci were clearly distinguished in two independent multiplex PCRs. Average minor allele frequency was 0.39, and accumulated matching probability was 2.12聽脳聽10^鈭?5. Complete profiles were successfully amplified with as little as 0.25聽ng of DNA. This method provides robust, sensitive, and cost-effective genotyping for human identification.