Developmental Validation of the MiSeq FGxTM Forensic Genomics System


Dr. Cydne Holt
Associate Director, Forensic Genomics
Illumina Inc.

Human DNA profiling using PCR of polymorphic short tandem repeat (STR) loci followed by capillary electrophoresis (CE) size separation and length-based allele typing has been the standard in the forensic community for over 20 years. Over the last decade, next-generation sequencing (NGS) has matured, bringing modern advantages to forensic DNA analysis.

The MiSeq FGx Forensic Genomics System, composed of the ForenSeq™ DNA Signature Prep Kit, MiSeq FGx Instrument, and ForenSeq Universal Analysis Software, simultaneously amplifies up to 231 forensic loci in a single multiplexed PCR. Targeted loci include 27 common forensic autosomal STRs, 24 Y-STRs, 7 X-STRs, amelogenin and three classes of single nucleotide polymorphism (SNPs). The System was subjected to developmental validation studies according to the 2012 Revised SWGDAM Validation Guidelines.

Approximately 1736 PCR reactions of the ForenSeq Kit were analyzed on the MiSeq FGx instrument using the ForenSeq software. Studies included but are not limited to species cross reactivity testing from a range of nonhuman organisms, DNA input sensitivity studies from 1 ng down to 7.8 pg, stochastic studies, two-person human DNA mixture testing with three genotype combinations, stability analysis using partially degraded DNA templates, and mock case type samples. Limitations of the MiSeq FGx System will be discussed. Results demonstrate that the MiSeq FGx System meets or exceeds forensic DNA quality assurance guidelines with robust, reliable, and reproducible performance on samples of various quantities and qualities. The presentation will include a presentation of the results form Dr. Cydne Holt, Associate Director of Forensic Genomics, and will conclude with a question and answer session.


For research, forensic, or paternity use only—not for clinical or therapeutic use in humans or animals.

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