The ForenSeq mtDNA Control Region Kit links a powerful assay with industry-leading sequencing and integrates seamlessly with intuitive software for optimum laboratory efficiency. The ForenSeq mtDNA Control Region workflow is completely flexible and scalable for all throughput requirements, allowing laboratories to quickly generate complete mtDNA control region results for up to 48 samples simultaneously in less than two working days. For low throughput requirements, the kit’s 48 reactions maintain a 12-month shelf-life and for higher demand laboratories, a similar library preparation routine makes automation easy.
- Short mean amplicon size for maximum variant detection
- Optimized for inhibitor tolerance
- Flexible and scalable throughput
- Integrated workflow for optimum laboratory efficiency
The high density of mutations concentrated in the mtDNA control region and the degraded nature of most forensic samples in which mtDNA analysis is used make optimal primer design challenging. The ForenSeq mtDNA Control Region kit is optimized to generate complete and actionable coverage, even for very low input amounts (see figure “The ForenSeq mtDNA Control Region system delivers extensive coverage across a wide range of DNA input amounts”).
To improve performance on highly degraded material, the ForenSeq mtDNA Control Region kit uses more than 120 primers, designed from recently curated mtDNA variant and frequency data, generating 18 primary amplicons in the mtDNA control region. All primary amplicons are <150 bp in length, many < 100 bp – the lowest average amplicon size of any current commercial mtDNA assay system. All amplicons have a minimum overlap of 3 bp to avoid gaps in sequence following post run, bioinformatic trimming. A buffer system optimized for inhibitors that creates a robust amplification environment (see figure “An improved ForenSeq buffer system maximizes performance“).
||1200 bp mtDNA control region
|DNA input recommendation
||100 pg gDNA and mtDNA per sample
||DNA extracted from a range of degraded samples, including bone, teeth and hair shaft
|Recommended multiplexing capacity
||3–48 libraries per run
|Number of primers
|Number of amplicons
|Average amplicon size
||≥ 3 bp
|Total library prep time
||7 hours and 45 minutes
||1 hour and 30 minutes