The introduction of massively parallel sequencing (MPS) in forensic investigation enables sequence-based large-scale multiplexing beyond size-based analysis using capillary electrophoresis (CE). For the practical application of MPS to forensic casework, many population studies have provided sequence data for autosomal short tandem repeats (STRs). However, SE33, a highly polymorphic STR marker, has little sequence-based data because of difficulties in analysis. In this study, 25 autosomal STRs were analyzed, including SE33, using an in-house MPS panel for 350 samples from four populations (African–American, Caucasian, Hispanic, and Korean). The barcoded MPS library was generated using a two-step PCR method and sequenced using a MiSeq System. As a result, 99.88% genotype concordance was obtained between length- and sequence-based analyses. In SE33, the most discordances (eight samples, 0.08%) were observed because of the 4 bp deletion between the CE and MPS primer binding sites. Compared with the length-based CE method, the number of alleles increased from 332 to 725 (2.18-fold) for 25 autosomal STRs in the sequence-based MPS method. Notably, additional 129 unique alleles, a 4.15-fold increase, were detected in SE33 by identifying sequence variations. This population data set provides sequence variations and sequence-based allele frequencies for 25 autosomal STRs.
Background—Single nucleotide polymorphisms (SNPs) have become popular in forensic genetics as an alternative to short tandem repeats (STRs) due to low mutation rates and small amplicon sizes. The Precision ID Identity Panel (Thermo Fisher Scientific), consisting of 90 autosomal SNPs and 34 Y-SNPs, was introduced for human identification by next-generation sequencing (NGS), enabling many studies on the global population; however, few reports are available on the Southeast Asian population.Methods and Results—A total of 96 unrelated male samples from Myanmar (Yangon) were analyzed with the Precision ID Identity Panel on a MiSeq (Illumina) using an in-house TruSeq compatible universal adapter. The sequencing performance was evaluated by locus balance and heterozygote balance, and the results were comparable to those of the Ion Torrent platform. For 90 autosomal SNPs, minor allele frequencies ranged between 0.068 and 0.500, and combined match probability (6.994×10−34) was lower than that of 22 PowerPlex Fusion autosomal STRs (3.130×10−26). Moreover, we identified 51 cryptic variations around the target SNPs using a custom variant caller, Visual SNP. For 34 Y-SNPs, 14 Y-haplogroups were observed—mostly O2 and O1b groups. Interpopulation analysis revealed that the Myanmar population is genetically closer to the East and Southeast Asian populations than the South Asian population.Conclusions—We demonstrate that the Precision ID Identity Panel can be successfully analyzed on a MiSeq using a custom data analysis pipeline and provide high discrimination power for human identification in the Myanmar population, while extending the accessibility of NGS analysis for SNPs in forensics.
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