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Genotyping of 38 insertion/deletion polymorphisms for human identification using universal fluorescent PCR.
- Source :
-
Molecular and cellular probes [Mol Cell Probes] 2014 Feb; Vol. 28 (1), pp. 13-8. Date of Electronic Publication: 2013 Sep 25. - Publication Year :
- 2014
-
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.<br /> (Copyright © 2013 Elsevier Ltd. All rights reserved.)
Details
- Language :
- English
- ISSN :
- 1096-1194
- Volume :
- 28
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Molecular and cellular probes
- Publication Type :
- Academic Journal
- Accession number :
- 24075877
- Full Text :
- https://doi.org/10.1016/j.mcp.2013.09.002