title: Forensic SNP genotyping with SNaPshot: Technical considerations for the development and optimization of multiplexed SNP assays.
abstract: This review explores the key factors that influence the optimization, routine use, and profile interpretation of the SNaPshot single-base extension (SBE) system applied to forensic single-nucleotide polymorphism (SNP) genotyping. Despite being a mainly complimentary DNA genotyping technique to routine STR profiling, use of SNaPshot is an important part of the development of SNP sets for a wide range of forensic applications with these markers, from genotyping highly degraded DNA with very short amplicons to the introduction of SNPs to ascertain the ancestry and physical characteristics of an unidentified contact trace donor. However, this technology, as resourceful as it is, displays several features that depart from the usual STR genotyping far enough to demand a certain degree of expertise from the forensic analyst before tackling the complex casework on which SNaPshot application provides an advantage. In order to provide the basis for developing such expertise, we cover in this paper the most challenging aspects of the SNaPshot technology, focusing on the steps taken to design primer sets, optimize the PCR and single-base extension chemistries, and the important features of the peak patterns observed in typical forensic SNP profiles using SNaPshot. With that purpose in mind, we provide guidelines and troubleshooting for multiplex-SNaPshot-oriented primer design and the resulting capillary electrophoresis (CE) profile interpretation (covering the most commonly observed artifacts and expected departures from the ideal conditions).
authors.affiliation: Forensic Genetics Unit, Department of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Galicia, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Fondevila
authors.affiliation: Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Børsting
authors.affiliation: Forensic Genetics Unit, Department of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Galicia, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Phillips
authors.affiliation: Forensic Genetics Unit, Department of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Galicia, Spain.
authors.forename: M
authors.initials: M
authors.lastname: de la Puente
authors.affiliation: http://www.euroforgen.eu/.
authors.forename: Euroforen-NoE
authors.initials: EN
authors.lastname: Consortium
authors.affiliation: Forensic Genetics Unit, Department of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Galicia, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Carracedo
authors.affiliation: Forensic Genetics Unit, Department of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Galicia, Spain.
authors.forename: N
authors.initials: N
authors.lastname: Morling
authors.affiliation: Forensic Genetics Unit, Department of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Galicia, Spain.
authors.forename: M V
authors.initials: MV
authors.lastname: Lareu
mesh_terms: D053803:Forensic Genetics / Q000379:methods*; D060005:Genotyping Techniques; D006801:Humans; D020641:Polymorphism, Single Nucleotide
keywords: Artifacts; CE; SBE; SNP; capillary electrophoresis; multiplex development; profile interpretation; single-base extension; single-nucleotide polymorphism