Use of Next‐Generation Sequencing in Forensic Genetics


The main purpose of forensic science institutions is to provide scientifically based investigations to the judicial system and assist society as a whole with objective analyses of evidence.

Ten years ago, the invention of clonal amplification by emulsion PCR or bridge PCR inspired a technical revolution in DNA sequencing commonly known as next‐generation sequencing (NGS). New methods and platforms were invented, all with the same purpose of sequencing massive numbers of DNA molecules simultaneously. These methods are of obvious interest to forensic genetic practitioners, who are frequently faced with the challenge of genotyping limited and degraded DNA material extracted from irreplaceable trace samples. NGS may be superior to the existing techniques and offers new exiting possibilities to the end users of forensic investigations.

Key Concepts

  • Next‐generation sequencing (NGS) is based on simultaneous clonal amplification of millions of individual DNA molecules and parallel sequencing of the generated products.
  • Fragment length analyses of PCR‐amplified short tandem repeats (STRs) have been the preferred method for human identification in forensic genetic case work for more than two decades.
  • The multiplexing capability of PCR‐NGS and sequencing capacity of NGS platforms make it possible to type different types of markers (STRs, SNPs and INDELs) in one assay, which increases the available information, saves time and reduces the overall cost of the investigation.
  • Human identification and forensic phenotyping may be accomplished simultaneously by typing human identification and phenotypical markers in one PCR‐NGS assay.
  • Detailed sequence information of STRs increases the statistical weight of the DNA evidence and may aid mixture interpretation.
  • Genome or transcriptome sequencing of samples from deceased makes it possible to combine forensic pathology with medical genetics and forensic toxicology with pharmacogenetics.

Keywords: next‐generation sequencing; massively parallel sequencing; PCR; forensic genetics; forensic genomics; human identification; human phenotyping; kinship testing; STR; SNP

Figure 1. An example of a DNA profile generated by PCR‐CE. The male individual was typed with the AmpFLSTR® NGM SElect™ PCR amplification Kit (ThermoFisher Scientific), and the PCR products were analysed by capillary electrophoresis. The electropherogram is divided into four images. Each image illustrates the detected PCR products carrying a specific fluorescent dye (blue, green, yellow (shown in black) and red). The abscissa indicates the length in nucleotides and the ordinate indicates the signal strength in relative fluorescent units. The expected size ranges of each locus are indicated by the grey boxes above the abscissa. The number of STR repeats for each product is indicated below the peak. ST = stutter. X and Y are the signals from the INDEL marker amelogenin. In the D10S1248 locus, the individual carry two alleles (13 and 15). Each of these alleles gave rise to a stutter (with a size of 12 and 14 repeats, respectively). In the D22S1045 locus, the individual is homozygous for allele 16 and two stutters are detected (with sizes of 15 and 17 repeats).
Figure 2. Three examples (blue, grey and brown eye colour) of digital eye images used for quantitative measurement of eye colour (Andersen et al., ). Among the pigmentary traits, eye colour has the highest heritability and phenotype predictability.


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Further Reading

Brion M, Sobrino B, Martinez M, et al. (2015) Massive parallel sequencing applied to the molecular autopsy in sudden cardiac death in the young. Forensic Science International: Genetics 18: 160–170.

Butler J (2005) Forensic DNA Typing: Biology, Technology and Genetics of STR Markers, 2nd edn. Burlington, MA: Elsevier Academic Press.

Butler J (2012) Advanced Topics in Forensic DNA Typing: Interpretation, 2nd edn. San Diego, CA: Elsevier Academic Press.

Børsting C and Morling N (2015) Next generation sequencing and its applications in forensic genetics. Forensic Science International: Genetics 18: 78–89.

Egeland T, Kling D and Mostad P (2016) Relationship Inference with Familias and R, 1st edn. San Diego, CA: Elsevier Academic Press.

Gill P (2014) Misleading DNA Evidence, 1st edn. Burlington, MA: Elsevier Academic Press.

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Phillips C, Gelabert‐Besada M, Fernandez‐Formoso L, et al. (2014) “New turns from old STaRs”: enhancing the capabilities of forensic short tandem repeat analysis. Electrophoresis 35: 3173–3187.

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Børsting, Claus, and Morling, Niels(Apr 2017) Use of Next‐Generation Sequencing in Forensic Genetics. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0027106]