title: Assay Development and Validation of an 8-SNP Multiplex Test to Predict Eye and Skin Coloration.
abstract: Identifying human remains is one of the many responsibilities of forensic scientists. An eye- and skin-color predictor translates genotypic information into phenotypic description. Eight single nucleotide polymorphisms (SNPs) are utilized for this predictor, five for eye, and six for skin coloration. Here, we describe the development and validation of an 8-SNP multiplex assay that consists of a multiplex PCR, followed by a multiplexed single-base primer extension reaction generating fluorescently labeled oligonucleotides of distinct length that are detected by multicolor capillary electrophoresis. Validation of this assay included tests for reproducibility, reliability, sensitivity, species specificity, its performance on degraded DNA, and on forensic samples. It can be concluded that the 8-SNP multiplex assay is robust and can be used on challenging samples, including bones, to reliably determine the genotypes to predict eye and skin color of individuals. This information can assist in the identification of human remains and missing persons.
authors.affiliation: Office of Chief Medical Examiner of the City of New York, Department of Forensic Biology, New York, NY.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Mushailov
authors.affiliation: Office of Chief Medical Examiner of the City of New York, Department of Forensic Biology, New York, NY.
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Rodriguez
authors.affiliation: Office of Chief Medical Examiner of the City of New York, Department of Forensic Biology, New York, NY.
authors.forename: Zoran M
authors.initials: ZM
authors.lastname: Budimlija
authors.forename: Mechthild
authors.initials: M
authors.lastname: Prinz
authors.affiliation: Office of Chief Medical Examiner of the City of New York, Department of Forensic Biology, New York, NY.
authors.forename: Elisa
authors.initials: E
authors.lastname: Wurmbach
mesh_terms: D000818:Animals; D053836:DNA Degradation, Necrotic; D019075:Electrophoresis, Capillary; D005127:Eye Color / Q000235:genetics*; D005453:Fluorescence; D053803:Forensic Genetics; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009841:Oligonucleotides / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012880:Skin Pigmentation / Q000235:genetics*; D013045:Species Specificity
chemical_list: D009841:Oligonucleotides
keywords: eye and skin color; forensic science; identification of human remains; multiplex assay; single nucleotide polymorphisms; validation