Figure 6.

Schematic of the mtDNA mutation assay. Sequences belonging to a given individual are grouped according to their multiplex identifier (MID) and according to their mtDNA primer identifier (PID). A consensus sequence is generated for each individual, based on all sequence groups containing at least five reads. Each group’s consensus sequence is then compared to the individuals’ consensus sequence. If a mutation (i.e., not present in the individual’ consensus sequence) is present in ≥75% of all reads belonging to a given group, it is categorized as a “true” mutation which is presumed to have been present in the original mtDNA molecule, at the time of primer labeling. All other mutations are deemed to represent PCR errors that occurred after the initial single‐cycle primer extension step or sequencing errors