A review of the method and validation of the MiSeq FGx™ Forensic Genomics Solution

The MiSeq FGx™ Forensic Genomics Solution (MFGS; Verogen Inc.) is a massively parallel sequencing workflow using Illumina sequencing technology. The workflow includes the ForenSeq™ DNA Signature Prep kit, the MiSeq FGx™, and the ForenSeq™ Universal Analysis Software (UAS). This review explores the molecular biology and bioinformatic methods used during the preparation of samples into sequencing libraries, the MiSeq FGx™ sequencing process, and the data analysis. We highlight what happens during each of these steps and how they influence the final results, while identifying limitations and possible improvements. After initial evaluations established its suitability for forensic applications, a number of studies have completed forensic validation studies. We review this work, and find the MFGS has shown to be a reliable and robust technology. The successful validation of the solution has allowed its implementation into forensic casework in a number of jurisdictions. Some questions still remain around the normalization of the sequencing libraries, and the bioinformatic processing and analysis of the sequencing results. In particular, the markers for biogeographical ancestry and externally visible characteristics require further research into their performance, and the ForenSeq™ UAS, in its current form, is limited in its ability to accurately predict these characteristics. We suggest a number of alternative bioinformatic tools that could be used instead. This article is categorized under: Forensic Biology > Forensic DNA Technologies Forensic Biology > Phenotypic Markers Forensic Biology > Ancestry Determination using DNA Methods