Figure 4. Assessment of major arc deletion detection resolution using a mixture series.

Two samples, one which harbors a major arc deletion at 61.5% heteroplasmy (mtDNA_Del) based on the multiplex quantification of the 1:0 neat sample, and one which does not harbor a known deletion (mtDNA_Normal), were mixed in varying proportions and analyzed in triplicate. The theoretical difference in mtDNA deletion ratio (mtDNA_DR) between each sample type is denoted with a triangle. Significant differences (α = 0.10) between the samples of varying deletion ratios are indicated with the brackets and asterisks; no significant difference was observed between the 1:0.75 mixture and the 1:1 mixture, the two mixtures which theoretically differ the least (mtDNA_DR difference of 4.4%). Therefore, this assay can detect differences in deletion ratio ≥ 5.9%.