In forensic biology, phenotype and ancestry estimations have received considerable attention due to their potential to provide investigative leads and aid in the human identification process. The IlluminaRTM MiSeq FGx(TM) in conjunction with the ForenSeq(TM) DNA Signature Prep kit is currently the only platform that produces genotypes of the CODIS-required STRs as well as provides phenotype and biogeographical ancestry estimations via phenotype-informative (piSNP) and ancestry-informative (aiSNP) markers, respectively. Although both markers have been developmentally validated for use in the field of forensic biology, there has been little data to determine the practical utility of these estimations to assist in the identification of missing persons using decedent casework samples.

This study investigated the accuracy and utility of both phenotypic and ancestral estimations based upon the DNA sequencing of 300 samples received by the Los Angeles County Department of Medical Examiner-Coroner (LACDoME-C). piSNP genotypes were translated into hair and eye colors using the Forenseq(TM) Universal Analysis Software (UAS) on the MiSeq FGx(TM) and the HIrisPlex System, and the statistical accuracy of these estimations was evaluated in context with the reported characteristics of the decedent. Similarly, estimations of each decedent's biogeographical ancestry, based upon the 56 aiSNPs, were compared to their reported ancestry to assess the efficacy of these markers in correctly predicting ancestry. This study reports on the capability of the FROG-kb database to adequately distinguish decedents beyond the Asian, African, European, and Admixed American global ancestries provided by the MiSeq FGx(TM) Universal Analysis Software's PCA plots. Specifically, FROG-kb was analyzed for its accuracy in correctly identifying individuals self-reported to possess Middle Eastern, Pacific Islander, Latin American, or Jewish ancestries.