1. Genotyping of 38 insertion/deletion polymorphisms for human identification using universal fluorescent PCR.
- Author
-
Oka K, Asari M, Omura T, Yoshida M, Maseda C, Yajima D, Matsubara K, Shiono H, Matsuda M, and Shimizu K
- Subjects
- Amelogenin genetics, DNA Primers, Fluorescent Dyes, Gene Frequency, Genetic Variation, Genotype, Humans, Japan, Sensitivity and Specificity, Forensic Anthropology methods, Genome, Human, Genotyping Techniques economics, INDEL Mutation, Multiplex Polymerase Chain Reaction, Polymorphism, Genetic
- Abstract
Short insertion/deletion (Indel) polymorphisms of approximately 2-6 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 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 × 10(-15). Complete profiles were successfully amplified with as little as 0.25 ng of DNA. This method provides robust, sensitive, and cost-effective genotyping for human identification., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF