Massively Parallel Sequencing (MPS), also known as Next Generation Sequencing (NGS), technologies have greatly enhanced the sensitivity, speed and informational power of DNA forensics. To turn massive amounts of sequencing data into usable forensic information, analysts rely on bioinformatics software. A new study of ExactID 2.0, a forensic NGS software tool from Battelle, demonstrates that it provides accurate genotypes using data generated by common commercially available forensic kits.
Battelle ExactID® 2.0 is the first commercially-available forensic NGS software tool that can be used to analyze data generated on a variety of NGS sequencing platforms. Unlike other commercial software programs, which are specific for the instrument they come with, ExactID can analyze data from many different types of NGS sequencers. That means that labs that have more than one type of sequencer can now analyze all of their data using a single software program and make apples-to-apples comparisons of data from multiple platforms.
ExactID allows users to access and evaluate the raw sequence data more thoroughly than platform-specific software tools. ExactID displays the sequence data so that stutter and noise patterns can be easily identified. Analytical threshold values can be set by the user and can be lowered to zero to allow for full evaluation and characterization of all sequence reads.
In order to ensure that ExactID provided accurate analysis for all types of sequencers, Battelle conducted a study using high-quality sequence data from three commercially available forensic kits generated on three different NGS platforms. The goal was to determine whether ExactID could generate accurate genotypes using data generated by each platform. Results from this study indicate that ExactID can be used to accurately evaluate forensic markers generated by any of the NGS platforms that currently support forensics applications.
For this experiment, eight samples were processed on the Thermo Fisher Personal Genome Machine (PGM), the Illumina MiSeq FGx and the MiSeq Desktop Sequencer. The HID AmpliSeq Ancestry and Identity single nucleotide polymorphism (SNP) panels were used on the PGM, the Illumina ForenSeq DNA signature prep kit was used for sequencing of SNPs and short tandem repeats on the MiSeq FGx, and the Promega PowerSeq kit was used to amplify autosomal and Y-chromosome short tandem repeat (STR) markers for sequencing on the MiSeq Desktop Sequencer instrument. All samples from all platforms were analyzed using the ExactID version 2.0 software.