title: The art of traditional native PAGE: The APLP 48-ID assay for human identification.
abstract: When full STR profiles cannot be obtained, further DNA analyses targeting single nucleotide polymorphisms (SNPs) may occasionally yield valuable information. Although the discrimination power of each SNP is relatively low, combined analysis of many SNPs can improve the personal identification ability to a level as high as that of commercial STR typing kits. In this study, we developed a new SNP typing method, named the amplified-product length polymorphism (APLP) 48-ID assay, for genotyping of 47 autosomal SNPs and two X and Y chromosomal markers for sex typing. Forty-seven SNPs were selected from all 22 autosomes, showing high diversity in European, Nigerian, Han Chinese, and Japanese population in the HapMap data. PCR primers were designed to generate amplicons 40-100 bp in length to increase the robustness of the PCR. The APLP 48-ID assay consisted of four independent PCR reactions followed by electrophoretic run on four lanes in a polyacrylamide gel. Complete profiles were obtained when more than 1.2 ng of DNA was used. We applied this assay for genotyping of 236 Japanese individuals. The random matching probability was 3.3E-20, and the power of exclusion was greater than 0.9999999. This method is a rapid, robust, and cost-effective approach for human identification and paternity testing.
authors.affiliation: Department of Forensic Medicine, Yamagata University School of Medicine, Yamagata 990-9585, Japan. Electronic address: kumetsu@med.id.yamagata-u.ac.jp.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Umetsu
authors.affiliation: Division of Legal Medicine, Faculty of Medicine, Tottori University, Yonago 683-8503, Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Yuasa
authors.affiliation: Department of Legal Medicine, Kansai Medical University, Hirakata 573-1010, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Hashiyada
authors.affiliation: Department of Legal Medicine, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Yamanashi 409-3898, Japan.
authors.forename: Noboru
authors.initials: N
authors.lastname: Adachi
authors.affiliation: Forensic Science Laboratory, Yamagata Prefectural Police Headquarters, Yamagata 990-8577, Japan.
authors.forename: Gotaro
authors.initials: G
authors.lastname: Watanabe
authors.affiliation: Department of Forensic Medicine, Yamagata University School of Medicine, Yamagata 990-9585, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Haneda
authors.affiliation: Department of Forensic Medicine, Yamagata University School of Medicine, Yamagata 990-9585, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yamazaki
mesh_terms: D044466:Asian People; D044383:Black People; D004247:DNA / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods*; D005260:Female; D053803:Forensic Genetics / Q000379:methods*; D005838:Genotype; D006801:Humans; D008297:Male; D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D044465:White People
chemical_list: D004247:DNA
keywords: Degraded DNA; Human identification; Multiplex amplified-product length polymorphism (APLP); SNPs; Short PCR products