Extract genetic data from small forensic samples

Forensic mtDNA Analysis with Sequencing

Illumina next-generation sequencing (NGS) has the potential to transform mitochondrial DNA (mtDNA) analysis. mtDNA analysis enables coroner and medical examiner laboratories to extract genetic data from small forensic samples found in less than ideal condition.

There are thousands of copies of mtDNA in every cell of the human body, but the copies found in different tissues may not be exactly the same. Referred to as heteroplasmy, these mitochondrial DNA mutations can make it difficult to confirm sample associations needed for identification in forensic investigations.

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Mitochondrial DNA Heteroplasmy

Dr. Mitch Holland of Penn State explains how high-throughput sequencing is helping us realize the power of mtDNA heteroplasmy in forensic research.

Illumina NGS technology, sequencing by synthesis (SBS), offers fast results and deep coverage of areas of interest. The MiSeq FGx System is capable of indexing 96 samples in a single sequencing run. Its simple workflow can quickly deliver forensic mtDNA analysis results, potentially speeding up investigations.

Additionally, the deep reads of Illumina NGS technology enable the accurate detection of mitochondrial DNA heteroplasmy with minor allele frequencies (MAFs) greater than 1%. This not only results in clearer heteroplasmy analysis, but also enables greater resolution of mixture samples.

mtDNA Sequencing AAFS Poster

Improved Human Mitochondrial DNA Analysis using Next-Generation Sequencing and Cloud-Based Computing and Storage.

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Following DNA extraction, mtDNA is amplified with a D-loop PCR protocol or with a whole mitochondrial genome approach. Prepare the mtDNA sequencing library with the Nextera XT DNA Library Prep Kit. Following library preparation, sequence mtDNA libraries on the MiSeq, or in the RUO mode of the MiSeq FGx.

Analyze your data using the MSR Plug-in and Variant Analyzer, or mtDNA Variant Processor and mtDNA Variant Analyzer BaseSpace Apps.

Click on the below to view products for each workflow step.

DNA Extraction and Quantification:
We recommend using your current DNA extraction and quantitation methods.


Library Preparation:
Nextera XT DNA Library Prep Kit (for mtDNA)

Prepare sequencing-ready libraries for small genomes like mitochondrial DNA in less than 90 minutes.

MiSeq FGx Instrument

First fully validated sequencing system for forensic genomics. Perform mtDNA sequencing in RUO mode.

MiSeq Reagent Kit v2

Sequencing reagents for mitochondrial DNA libraries.

mtDNA Variant Processor BaseSpace App

Enables streamlined variant analysis of d-loop and whole mitochondrial DNA sequence data.

mtDNA Variant Analyzer BaseSpace App

Enables simplified visualization of d-loop and whole mitochondrial DNA sequence data.

Identifying Missing Persons
mtDNA Analysis to Identify Missing Persons

NGS delivers a single platform and workflow to overcome the wide range of difficult samples encountered in missing persons cases. Learn More »

Disaster Victim Identification
Disaster Victim Identification

Genetic analysis of mass fatality evidence with NGS lets you extract as much information as possible from highly compromised samples.
Learn More »

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Human mtDNA Analysis in BaseSpace Sequence Hub

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