The mitochondrial genome is a circular DNA molecule which is distinct from the nuclear genome. In humans, it’s about 16 kb long and encodes 37 genes. Mitochondrial disorders arise as a result of dysfunction of the mitochondrial respiratory chain, and often times, these are caused by mutations of genes encoded by mitochondrial DNA (mtDNA). Because there is considerable clinical variability between mitochondrial disorders and many patients who exhibit phenotypes that overlap diseases, often diagnosis may only be confirmed by identification of a pathogenic mtDNA variant through molecular genetic testing of DNA extracted from a blood sample.

DescriptionThe assay kit contains all the reagents required for capture of the entire human mitochondrial DNA genome and library preparation for next-gen sequencing
Capture MethodThe VariantPro™ system incorporates novel Relay-PCR™ and Omega Primer™ technologies to form a one-step multiplex PCR based workflow.
Amplicon Coverage
Designed for 100% amplicon coverage of all regions of the mitochondrial genome (16,569 bp)
Amplicon Length Average 200 bp
Primer Pools
110 pairs of primers in 2 primer pair pools
Input DNA Required 1-5 ng human genomic DNA per reaction tube (2) – depending on the DNA quality

Assay Time4-6 hours
Hands-on Time5 minutes
Sequencing PlatformIllumina
Multiplexing CapabilityUp to 3024 samples in one lane
Coverage Uniformity> 99% at 0.2 x mean
Dropout Rate0

Capture Technology

VariantPro targeted sequencing system is based on a novel multiplex PCR technology that combines major innovations to facilitate simple operation.…

Amplicon List

Refer to User Guide – VariantPro Amplicon Design Files for detail of the amplicon design scheme. geneName: MT-CO1;MT-TS1;MT-TD;MT-CO2…

Workflow

In a single tube, a pair of common primers are added into a mixture of multiple pairs of specific primers and genomic DNA to form a single step, two stage multiplex PCR reaction….

uniformity

mtDNA-allele

Coverage uniformity: > 99.1 % of amplicons detected at > 0.2X mean coverage. The lowest and highest covered amplicons varied within only 1.35 log at 2500 read depth

The detected mutation frequencies matched closely to the actual mixing frequencies of spike-in variant genome.

More performance data can be found here – Technical Note – Performance of the VariantPro Mitochondrial Panel