Research Article| Volume 63, 102819, March 2023

Download started.


A global snapshot of current opinions of next-generation sequencing technologies usage in forensics

Published:December 09, 2022DOI:


      • General knowledge and experience to NGS applies to forensic laboratories worldwide.
      • Limited funding and training have the largest impact on NGS implementation plans.
      • Main concerns include lack of bioinformatics support and statistical applications.
      • In the 1–5 years, CE will remain the main technology, with an increase of NGS.
      • A gradual CE to NGS technology shift is expected in 5–10 + years.


      The future of forensic DNA testing is being shaped by the research and usage of next-generation systems, which have increased the multiplexing capabilities of the field and the type and amount of genetic data that can be utilized for investigations. The NGS adoption for casework has been slow, albeit the plethora of data that has been published. This study evaluated the current opinions on sequencing in forensics. A 20-question online-survey focusing on NGS knowledge, training, and usage was distributed to 6001 forensic DNA researchers and practitioners worldwide. A total of 367 responses were obtained from all continents (North/South America (69.8%), Europe (21.2%), Asia (5.5%), Oceania (2.5%), and Africa (1%)). The respondents consisted of 50% practitioners, 31% researchers, and 19% both. Of these, 38% already own a next-gen sequencing instrument, and 13% are planning to purchase one. Overall, there exists an extensive knowledge on next-gen sequencing within the forensic community, including among laboratories that have not yet implemented this high-throughput technology in their workflows. Current usage focuses primarily on SNP analysis for investigative leads and mitochondrial DNA analysis while future applications included both STR and SNP testing applied to general casework. The major overall concerns respondents have for implementing a sequencing instrument include limited funding, staffing, lack of time, and the cost-effectiveness of providing this service. Specific technical concerns that the respondents had are the lack of training, statistical applications, bioinformatics support, and of rigorous guidelines and recommendations. Most of the respondents do believe there will be a technology shift from using CE only to the use of NGS on casework in 5–10 years. In addition, around 66% of respondents believe that it is moderately to very likely that the court will accept sequencing analysis. Sixteen percent fell in the middle, and the remaining 15% believe it is more unlikely, with 3% of respondents believing it is very unlikely. In conclusion, this work outlines current analytical challenges experienced by the global forensic DNA community and addresses different strategies for the implementation of next-gen sequencing technologies in casework.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Forensic Science International: Genetics
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Bulter J.M.
        Advanced Topics in Forensic DNA Typing: Methodology.
        Elsevier, Waltham, MA2012
        • Haddrill P.R.
        Developments in forensic dna analysis.
        Emerg. Top. Life Sci. 2021; 5: 381-393
        • Oldoni F.
        • Podini D.
        Forensic molecular biomarkers for mixture analysis.
        Forensic Sci. Int. Genet. 2019; 41: 107-119
        • Jesus Alvarez-Cubero M.
        • Saiz M.
        • Martínez-García B.
        • et al.
        Next generation sequencing: an application in forensic sciences.
        Ann. Hum. Biol. 2017; 44: 581-592
        • Børsting C.
        • Morling N.
        Next generation sequencing and its applications in forensic genetics.
        Forensic Sci. Int. Genet. 2015; 18: 78-89
        • Ballard D.
        • Winkler-Galicki J.
        • Wesoły J.
        Massive parallel sequencing in forensics: advantages, issues, technicalities, and prospects.
        Int. J. Leg. Med. 2020; 134: 1291-1303
        • de Knijff P.
        From next generation sequencing to now generation sequencing in forensics.
        Forensic Sci. Int. Genet. 2019; 38: 175-180
        • Köcher S.
        • Müller P.
        • Berger B.
        • et al.
        Inter-laboratory validation study of the ForenSeqTM DNA Signature Prep Kit.
        Forensic Sci. Int. Genet. 2018; 36: 77-85
        • Brandhagen M.D.
        • Just R.S.
        • Irwin J.A.
        Validation of NGS for mitochondrial DNA casework at the FBI Laboratory.
        Forensic Sci. Int. Genet. 2020; 44 (Article)102151
        • Walsh S.
        • Wollstein A.
        • Liu F.
        • et al.
        DNA-based eye colour prediction across Europe with the IrisPlex system.
        Forensic Sci. Int. Genet. 2012; 6: 330-340
        • Hussing C.
        • Huber C.
        • Bytyci R.
        • et al.
        Sequencing of 231 forensic genetic markers using the MiSeq FGx TM forensic genomics system-an evaluation of the assay and software.
        Forensic Sci. Res. 2018; 3: 111-123
        • Zeng X.
        • King J.
        • Hermanson S.
        • et al.
        An evaluation of the PowerSeqTM Auto System: a multiplex short tandem repeat marker kit compatible with massively parallel sequencing.
        Forensic Sci. Int. Genet. 2015; 19: 172-179
        • Faccinetto C.
        • Serventi P.
        • Staiti N.
        • et al.
        Internal validation study of the next generation sequencing of GlobalfilerTM PCR amplification kit for the Ion Torrent S5 sequencer.
        Forensic Sci. Int. Genet. Suppl. Ser. 2019; 7: 336-338
        • Hollard C.
        • Ausset L.
        • Chantrel Y.
        • et al.
        Automation and developmental validation of the ForenSeqTM DNA Signature Preparation kit for high-throughput analysis in forensic laboratories.
        Forensic Sci. Int. Genet. 2019; 40: 37-45
        • Guo F.
        • Yu J.
        • Zhang L.
        • Li J.
        Massively parallel sequencing of forensic STRs and SNPs using the Illumina® ForenSeqTM DNA Signature Prep Kit on the MiSeq FGxTM Forensic Genomics System.
        Forensic Sci. Int. Genet. 2017; 31: 135-148
        • Sharma V.
        • van der Plaat D.A.
        • Liu Y.
        • Wurmbach E.
        Analyzing degraded DNA and challenging samples using the ForenSeqTM DNA Signature Prep kit.
        Sci. Justice. 2020; 60: 243-252
        • Elwick K.
        • Bus M.M.
        • King J.L.
        • et al.
        Utility of the Ion S5TM and MiSeq FGxTM sequencing platforms to characterize challenging human remains.
        Leg. Med. 2019; 41 (Article)101623
        • Montano E.A.
        • Bush J.M.
        • Garver A.M.
        • et al.
        Optimization of the Promega PowerSeqTM Auto/Y system for efficient integration within a forensic DNA laboratory.
        Forensic Sci. Int. Genet. 2018; 32: 26-32
        • Bruijns B.
        • Tiggelaar R.
        • Gardeniers H.
        Massively parallel sequencing techniques for forensics: a review.
        Electrophoresis. 2018; 3: 2642-2654
        • Heidegger A.
        • Xavier C.
        • Niederstätter H.
        • et al.
        Development and optimization of the VISAGE basic prototype tool for forensic age estimation.
        Forensic Sci. Int. Genet. 2020; 48 (Article)102322
        • Marano L.A.
        • Fridman C.
        DNA phenotyping: current application in forensic science.
        Res. Rep. Forensic Med. Sci. 2019; 9: 1-8
        • Schneider P.M.
        • Prainsack B.
        • Kayser M.
        The use of forensic DNA phenotyping in predicting appearance and biogeographic ancestry.
        Dtsch. Arztebl. Int. 2019; 116: 873-880
        • Kling D.
        • Phillips C.
        • Kennett D.
        • Tillmar A.
        Investigative genetic genealogy: current methods, knowledge and practice.
        Forensic Sci. Int. Genet. 2021; 52 (Article)102474
        • Oldoni F.
        • Kidd K.K.
        • Podini D.
        Microhaplotypes in forensic genetics.
        Forensic Sci. Int. Genet. 2019; 38: 54-69
        • El-Mogy M.
        • Lam B.
        • Haj-Ahmad T.A.
        • et al.
        Diversity and signature of small RNA in different bodily fluids using next generation sequencing.
        BMC Genom. 2018; 19 (Article)418
        • Vidaki A.
        • Daniel B.
        • Court D.S.
        Forensic DNA methylation profiling—potential opportunities and challenges.
        Forensic Sci. Int. Genet. 2013; 7: 499-507
        • Greytak E.M.
        • Moore C.C.
        • Armentrout S.L.
        Genetic genealogy for cold case and active investigations.
        Forensic Sci. Int. Genet. 2019; 299: 103-113
        • Faccinetto C.
        • Sabbatini D.
        • Serventi P.
        • et al.
        Internal validation and improvement of mitochondrial genome sequencing using the Precision ID mtDNA Whole Genome Panel.
        Int. J. Leg. Med. 2021; 135: 2295-2306
        • Zhou Y.
        • Guo F.
        • Yu J.
        • et al.
        Strategies for complete mitochondrial genome sequencing on Ion Torrent PGMTM platform in forensic sciences.
        Forensic Sci. Int. Genet. 2016; 22: 11-21
        • National Research Council
        Strengthening Forensic Science in the United States: A Path Forward.
        The National Academies Press, Washington, D.C.2009
      1. Recommendations of the DNA Commission. Available at:, 2022 (accessed August 17, 2022).

      2. Scientific Working Group on DNA Analysis Methods, SGWDAM Validation Guidelines for DNA Analysis Methods (2016). Available at:, (accessed 16 August 2022).

      3. Federal Bureau of Investigation, Quality Assurance Standards for Forensic DNA Testing Laboratories (2020). Available at:, (accessed 16 Aug 2022).

        • Alonso A.
        • Müller P.
        • Roewer L.
        • et al.
        European survey on forensic applications of massively parallel sequencing.
        Forensic Sci. Int. Genet. 2017; 29: E23-E25
        • Gross T.E.
        • Fleckhaus J.
        • Schneider P.M.
        Progress in the implementation of massively parallel sequencing for forensic genetics: results of a European-wide survey among professional users.
        Int. J. Leg. Med. 2021; 135: 1425-1432
      4. Qualtrics® (2022). Available at:, (accessed 16 Aug 2022).

      5. The International Society of Forensic Genetics (2022). Available at:, (accessed 16 Aug 2022).

      6. American Academy of Forensic Science (2022). Available at:, (accessed 16 Aug 2022).

        • Dale W.M.
        • Becker W.S.
        Forensic Laboratory Management.
        CRC Press, Boca Raton, FL2015
        • Parson W.
        • Ballard D.
        • Budowle B.
        • et al.
        Massively parallel sequencing of forensic STRs: considerations of the DNA commission of the International Society for Forensic Genetics (ISFG) on minimal nomenclature requirements.
        Forensic Sci. Int. Genet. 2016; 22: 54-63
        • Gettings K.B.
        • Ballard D.
        • Bodner M.
        • et al.
        Report from the STRAND Working Group on the 2019 STR sequence nomenclature meeting.
        Forensic Sci. Int. Genet. 2019; 43 (Article)102165
      7. Scientific Working Group on DNA Analysis Methods, SWGDAM Interpretation Guidelines for Mitochondrial DNA Analysis by Forensic DNA Testing Laboratories (2019). Available at:, (accessed 16 Aug 2022).

      8. Scientific Working Group on DNA Analysis Methods, Addendum to “SWGDAM Interpretation Guidelines for Autosomal STR Typing by Forensic DNA Testing Laboratories” to Address Next Generation Sequencing (2019). Available at:, (accessed 16 Aug 2022).

      9. OSAC Update Meeting Bio Sac (2019). Available at:, (accessed 16 Aug 2022).