Experimental Strategy of Mitochondrial Genomics

The vertebrate mitochondrial (mtDNA) genome is a circular 17Kbp molecule that comprises 13 protein-coding regions, two rRNA genes, a replication control region, and 22 tRNA genes [top left]. A PCR-primer design strategy based on conserved sequences in closely-related species generates the complete mtDNA sequence as a series of overlapping, contiguous gene sequences (“contigs”) [bottom left]. A series of forward (red boxes) and reverse (blue boxes) PCR primers amplify the entire mtDNA molecule as an ordered series of 20 ~ 30 overlapping contigs (red- & blue-tipped boxes).