Material recovered will be used as a template for mitochondrial genome discovery.
Because mitochondrial DNA comprises only about 2% of Vigna total DNA [49], the effective concentration of mitochondrial template molecules in PCR carried out using purified mitochondrial DNA was about 70 times that using total DNA.
We also observed greater variance associated with sequence coverage among samples for the mitochondrial genome, which could reflect a difference in the amount of mitochondrial template present in each DNA sample (Additional file 3: Figure S4; Additional file 5).
This interpretation is consistent with results reported for a cnidarian, the jellyfish A. aurita (de Paula et al. 2013), and might therefore be predicted to apply as a basic pattern of template mitochondrial transmission within the entire animal kingdom.
However, in contrast to the hemizygous genomic loci, which were known to be present at a single copy in the template DNA, the copy number of mitochondrial genomes was not known in either the single cell samples or in the tail clipping DNA that was used for the dilution series.
The native template to competitive template ratio was calculated and the number of mitochondrial or nuclear DNA molecules was estimated according to Willey et al. [ 7].
Studies that investigate old and degraded DNA have as a rule demonstrated that the inherent high ratio of mitochondrial to nuclear template molecules will result in mtDNA remaining PCR-amplifiable for a longer period of time than nuDNA in any specific sample [3].
This is also supported by PCR amplifications of the recombinant configurations from Vigna total DNA, as these reactions contained only about 1/70th the level of mitochondrial genomes (see Methods) as the purified mitochondrial DNA template used in the assays described at the beginning of this section.
Since in DUI species sperm mitochondria are inherited through male lineages, the need for M mitochondria to function both as energy supply for sperm swimming and as genetic templates represents an ideal situation to study the relationship between mitochondrial activity, mtDNA damage and faithful transmission of mitochondrial genetic information.
Avery was given a diagnosis of mitochondrial disorder at birth.
