[Single-fiber polymerase chain reaction for detection of mutant mitochondrial DNA].
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.
Tokunaga
M
M
Department of Neurology, Kumamoto University.
Mita
S
S
Uchino
M
M
D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics*; D006801:Humans; D017237:Mitochondrial Encephalomyopathies / Q000235:genetics; D018485:Muscle Fibers, Skeletal / Q000737:chemistry; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods
D004272:DNA, Mitochondrial