International Journal of Legal Medicine

, Volume 133, Issue 2, pp 325–334 | Cite as

The Danish STR sequence database: duplicate typing of 363 Danes with the ForenSeq™ DNA Signature Prep Kit

  • C. Hussing
  • R. Bytyci
  • C. Huber
  • N. Morling
  • C. BørstingEmail author
Original Article


Some STR loci have internal sequence variations, which are not revealed by the standard STR typing methods used in forensic genetics (PCR and fragment length analysis by capillary electrophoresis (CE)). Typing of STRs with next-generation sequencing (NGS) uncovers the sequence variation in the repeat region and in the flanking regions. In this study, 363 Danish individuals were typed for 56 STRs (26 autosomal STRs, 24 Y-STRs, and 6 X-STRs) using the ForenSeq™ DNA Signature Prep Kit to establish a Danish STR sequence database. Increased allelic diversity was observed in 34 STRs by the PCR-NGS assay. The largest increases were found in DYS389II and D12S391, where the numbers of sequenced alleles were around four times larger than the numbers of alleles determined by repeat length alone. Thirteen SNPs and one InDel were identified in the flanking regions of 12 STRs. Furthermore, 36 single positions and five longer stretches in the STR flanking regions were found to have dubious genotyping quality. The combined match probability of the 26 autosomal STRs was 10,000 times larger using the PCR-NGS assay than by using PCR-CE. The typical paternity indices for trios and duos were 500 and 100 times larger, respectively, than those obtained with PCR-CE. The assay also amplified 94 SNPs selected for human identification. Eleven of these loci were not in Hardy-Weinberg equilibrium in the Danish population, most likely because the minimum threshold for allele calling (30 reads) in the ForenSeq™ Universal Analysis Software was too low and frequent allele dropouts were not detected.


Short tandem repeats Danes ForenSeq™ Forensic genetics Next-generation sequencing 



The authors thank Anja Ladegaard Jørgensen for technical support, and Carina Grøntved Jønck and Brian Stidsen for bioinformatics support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the national research committee (the Danish ethical committee, H-1-2011-081) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Samples were taken from the biobank of the Department of Forensic Medicine, University of Copenhagen (RIBVF; approved by the Danish Data Protection Agency, 2002-54-1080). The Danish ethical committee waived the requirement for informed consent (H-4-2011-081). This article does not contain any studies with animals performed by any of the authors.

Supplementary material

414_2018_1854_MOESM1_ESM.pdf (627 kb)
ESM 1 (PDF 626 kb)
414_2018_1854_MOESM2_ESM.xlsx (47 kb)
ESM 2 (XLSX 47 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • C. Hussing
    • 1
  • R. Bytyci
    • 1
  • C. Huber
    • 1
  • N. Morling
    • 1
  • C. Børsting
    • 1
    Email author
  1. 1.Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical SciencesUniversity of CopenhagenCopenhagenDenmark

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