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        Analysis of Mitochondrial DNA Mutations: Point Mutations

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        Since the first demonstration that mutations of the mitochondrial genome were associated with human disease, more than 100 pathological mitochondrial DNA (mtDNA) defects have been characterized in patients with a broad spectrum of clinical manifestations (1 ). Single-point mutations, involving either protein-encoding genes or more commonly RNA (rRNA and tRNA) genes, represent a substantial proportion (more than one third) of the pathogenic mtDNA mutations described in the literature, and this number is steadily increasing (2 , 3 ). Although some of the more common mtDNA point mutations can be screened using simple polymerase chain reaction (PCR)-based techniques (e.g., restriction digest analysis), an increasing number of pathological point mutations are identified only when large-scale sequencing of either all 22 tRNA genes or the whole mitochondrial genome is performed (4 7 ).
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