title: Single nucleotide polymorphism typing with massively parallel sequencing for human identification.
abstract: The Ion AmpliSeq™ HID single nucleotide polymorphism (SNP) panel, a primer pool of 103 autosomal SNPs and 33 Y-SNPs, was evaluated using the Ion 314™ Chip on the Ion PGM™ Sequencer with four DNA samples. The study focused on the sequencing of DNA at three different initial target quantities, related interpretation issues, and concordance of results with another sequencing platform, i.e., Genome Analyzer IIx. With 10 ng of template DNA, all genotypes at the 136 SNPs were detected. With 1 ng of DNA, all SNPs were detected and one SNP locus in one sample showed extreme heterozygote imbalance on allele coverage. With 100 pg of DNA, an average of 1.6 SNP loci were not detected, and an average of 4.3 SNPs showed heterozygote imbalance. The average sequence coverage was 945-600× at autosomal SNPs and 465-209× at Y-SNPs for 10 ng-100 pg of DNA. The average heterozygote allele coverage ratio was 89.6-61.8 % for 10 ng-100 pg of DNA. At 10 ng of DNA, all genotypes of the 95 SNPs shared between the two different sequencing platforms were concordant except for one SNP, rs1029047. The error was due to the misalignment of a flanking homopolymer. Overall, the data support that genotyping a large battery of SNPs is feasible with massively parallel sequencing. With barcode systems, better allele balance, and specifically designed alignment software, a more comprehensive rapid genotyping and more cost-effective results may be obtained from multiple samples in one analysis than are possible with current typing and capillary electrophoresis systems.
authors.affiliation: Institute of Applied Genetics, Department of Forensic and Investigative Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Fort Worth, TX, 76107, USA.
authors.forename: Seung Bum
authors.initials: SB
authors.lastname: Seo
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: King
authors.forename: David H
authors.initials: DH
authors.lastname: Warshauer
authors.forename: Carey P
authors.initials: CP
authors.lastname: Davis
authors.forename: Jianye
authors.initials: J
authors.lastname: Ge
authors.forename: Bruce
authors.initials: B
authors.lastname: Budowle
mesh_terms: D000483:Alleles; D004247:DNA / Q000235:genetics*; D005240:Feasibility Studies; D005260:Female; D053803:Forensic Genetics / Q000295:instrumentation / Q000379:methods*; D005784:Gene Amplification; D015723:Gene Library; D006580:Genetic Carrier Screening; D056426:Genetic Loci / Q000235:genetics; D005838:Genotype; D006801:Humans; D008297:Male; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation
chemical_list: D004247:DNA