Single-fiber polymerase chain reaction for detection of mutant mitochondrial DNA] |
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Authors: | M Tokunaga S Mita M Uchino |
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Affiliation: | Department of Neurology, Kumamoto University. |
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Abstract: | Single-fiber PCR amplifies mitochondrial DNA (mtDNA) in single muscle fiber isolated from cross frozen section. The PCR products are digested with a restriction enzyme to distinguish mutant mtDNA from wild-type mtDNA. The proportion of mutant mtDNA is higher in ragged-red fiber (RRF) than in non-RRF in mitochondrial encephalomyopathies with mutations of mtDNA. This method may be applied to evaluate amount of mtDNA and mRNA in single muscle fiber, and become a powerful tool to elucidate the pathogenetic mechanism in mitochondrial encephalomyopathies. |
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