Forensic DNA Analysis: Massively Parallel Sequencing Workflows
Massively parallel sequencing (MPS) offers a solution to the biggest challenges facing CE methods, such as distinguishing true allele variation from PCR artifacts, interpreting mixtures, and obtaining usable profiles from degraded DNA. However, adopting a new technology can have unique challenges. The PowerSeq® product family can help make the transition a smooth one.
PowerSeq® products enable target amplification of the mitochondrial control region and whole mitochondrial sequences. In addition, PowerSeq® products can amplify short tandem repeats (STRs) in both Y-chromosome and autosomal DNA to prepare Illumina® sequencing libraries for MPS analysis.
PowerSeq® Whole Mito System
Streamlined workflow for complete mtDNA analysis
The PowerSeq® Whole Mito System generates 161 small amplicons covering the entire region of the mitochondrial genome. The targeted regions for amplification are designed for an average size of 167bp to ensure optimal results from degraded samples. The system contains reagents for library preparation and a streamlined workflow that greatly reduces the number of steps and time required to produce libraries ready for sequencing.
PowerSeq® CRM Nested System, Custom
Streamlined library prep for mtDNA analysis
The PowerSeq® CRM Nested System, Custom, generates 10 small amplicons covering the control region of the mitochondrial genome in one multiplex reaction. This technique improves the results for degraded samples. The workflow greatly simplifies library preparation by saving time, decreasing sample loss and reducing data variability.
PowerSeq® 46GY System
Optimized panel of loci for STR analysis
The PowerSeq® 46GY System offers the same sensitivity as STR amplification kits. It can be used to prepare MPS libraries and generate sequencing data compatible with Illumina® sequencing technology. The kit enables identification of sequence variants in familiar autosomal and Y-STR loci, so that data generated are usable and interpretable with current guidelines.
PowerSeq® Quant MS System
Consistent library prep for STR analysis
The PowerSeq® Quant MS System is a qPCR-based system designed for quantitation of libraries prior to sequencing on the Illumina® MiSeq® system. Accurate library quantitation provides balanced representation of pooled libraries and reduces sequencing bias.
Massively Parallel Sequencing Product Groups
Preprocessing and Differential Extraction
Reagents to help you pretreat swabs and storage card punches prior to direct amplification, preprocess challenging casework samples and easily separate sperm and epithelial cells.
DNA Isolation
DNA extraction systems that allow you to yield pure, intact DNA and save time with automation.
Human Specific DNA Quantitation
Systems to help you determine human genomic DNA concentration prior to STR amplification, assess DNA integrity and detect PCR inhibitors.
Target Amplification and Library Prep
Reagents to facilitate library prep and amplification of the mitochondrial control region.
Massively Parallel Sequencing for Forensic DNA Analysis
Traditionally, capillary electrophoresis (CE) has been the method of choice for analyzing short tandem repeats (STRs) for human identification. Although CE methods continue to evolve in terms of increasing sensitivity and the ability to work with degraded or poor-quality DNA, the technique still faces some challenges.
Next-generation sequencing (NGS), or massively parallel sequencing (MPS), enables the simultaneous analysis of hundreds of genetic markers, considerably more than current CE technologies. In addition to providing information on the size of the repeated regions, as CE does, MPS determines the underlying DNA sequence of each region. In doing so, MPS offers a solution to the biggest challenges facing CE methods, such as distinguishing true allele variation from PCR artifacts, interpreting mixtures, and obtaining usable profiles from degraded DNA.
A popular application of MPS in forensics is the sequencing of mitochondrial DNA—either the variable control region or the whole mitochondrial genome—with increased mixture deconvolution and heteroplasmy resolution compared to traditional methods. Although adoption of MPS workflows by forensic laboratories has been slow, the technology holds great promise for increasing the accuracy and throughput of forensic DNA analysis. Future advancements in MPS to further lower the cost will encourage more widespread adoption and consequent validation of forensic MPS methods.
