title: A SNP panel for identity and kinship testing using massive parallel sequencing.
abstract: Within forensic genetics, there is still a need for supplementary DNA marker typing in order to increase the power to solve cases for both identity testing and complex kinship issues. One major disadvantage with current capillary electrophoresis (CE) methods is the limitation in DNA marker multiplex capability. By utilizing massive parallel sequencing (MPS) technology, this capability can, however, be increased. We have designed a customized GeneRead DNASeq SNP panel (Qiagen) of 140 previously published autosomal forensically relevant identity SNPs for analysis using MPS. One single amplification step was followed by library preparation using the GeneRead Library Prep workflow (Qiagen). The sequencing was performed on a MiSeq System (Illumina), and the bioinformatic analyses were done using the software Biomedical Genomics Workbench (CLC Bio, Qiagen). Forty-nine individuals from a Swedish population were genotyped in order to establish genotype frequencies and to evaluate the performance of the assay. The analyses showed to have a balanced coverage among the included loci, and the heterozygous balance showed to have less than 0.5 % outliers. Analyses of dilution series of the 2800M Control DNA gave reproducible results down to 0.2 ng DNA input. In addition, typing of FTA samples and bone samples was performed with promising results. Further studies and optimizations are, however, required for a more detailed evaluation of the performance of degraded and PCR-inhibited forensic samples. In summary, the assay offers a straightforward sample-to-genotype workflow and could be useful to gain information in forensic casework, for both identity testing and in order to solve complex kinship issues.
authors.affiliation: Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Artillerigatan 12, SE-58758, Linköping, Sweden.
authors.forename: Ida
authors.initials: I
authors.lastname: Grandell
authors.affiliation: QIAGEN Sciences, Inc, 6951 Executive Way, Frederick, MD, 21703, USA.
authors.forename: Raed
authors.initials: R
authors.lastname: Samara
authors.affiliation: Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Artillerigatan 12, SE-58758, Linköping, Sweden. andreas.tillmar@rmv.se.|Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, Sandbäcksgatan 7, SE-582 35, Linköping, Sweden. andreas.tillmar@rmv.se.
authors.forename: Andreas O
authors.initials: AO
authors.lastname: Tillmar
mesh_terms: D016172:DNA Fingerprinting; D005787:Gene Frequency; D005838:Genotype; D006579:Heterozygote; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D010375:Pedigree; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D013548:Sweden
keywords: Forensic genetics; Human identification; Kinship; Next generation sequencing; Single-nucleotide polymorphism