The ForenSeq mtDNA Whole Genome Kit features the highest resolution for mtGenome sequencing in the Verogen library prep portfolio. Tiled amplicons, fit-for-purpose buffers, and a thoughtfully designed protocol enable support for the lower end of DNA input amounts, various sample types, and a broad range of forensic and database applications, including missing persons, disaster victim identification (DVI), and population studies.
- High sensitivity across a range of gDNA inputs
- Enhanced recovery of low input samples
- Robust resistance to common inhibitors
The protocol includes an optional second purification to further improve the performance of low-level and complex samples. The figure “High coverage across a range of input DNA amounts” presents a comparison of coverage results for libraries with and without the second purification. Compared to a single purification, two purifications show progressive coverage advancements, culminating with 90% improvement at 2 pg (see figure “Read count improvement with a second purification”). Another challenge of forensic sample analysis is the frequent presence of PCR inhibitors that can compromise library prep. When varying amounts of three inhibitors common in forensic mtDNA samples were added to 100 pg of Control DNA HL60, results shown in the figure “Buffers highly resistant to relevant PCR inhibitors” demonstrated high resistance with consistent coverage across all inputs.
|DNA input recommendation
||100 pg gDNA per sample
||16 libraries per run
|Number of primers
|Number of amplicons
|Average amplicon size
|Total library prep time
||7 hours and 45 minutes
||1 hour and 45 minutes