title: Collection of isolated cells for studying mitochondrial DNA mutations within individual cells.
abstract: Mitochondrial genome integrity is an important issue in somatic mitochondrial genetics. Development of quantitative methods is indispensable to somatic mitochondrial genetics as quantitative studies are required to characterize heteroplasmy and mutation processes, as well as their effects on phenotypic developments. This chapter outlines the methods for collecting individual cells appropriate for analysis of mtDNA mutations by single-molecule PCR. In addition, we describe the protocols for respiratory complexes II and IV in these cells. Together, the identification of respiratory deficiency and mtDNA mutations within single cells provides a powerful means of evaluating the importance of these events in disease and aging.
authors.affiliation: Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, USA.
authors.forename: Yevgenya
authors.initials: Y
authors.lastname: Kraytsberg
authors.forename: Natalya
authors.initials: N
authors.lastname: Bodyak
authors.forename: Susan
authors.initials: S
authors.lastname: Myerow
authors.forename: Alexander
authors.initials: A
authors.lastname: Nicholas
authors.forename: Konstantin
authors.initials: K
authors.lastname: Ebralidze
authors.forename: Konstantin
authors.initials: K
authors.lastname: Khrapko
mesh_terms: D004252:DNA Mutational Analysis; D004272:DNA, Mitochondrial / Q000235:genetics*; D042963:Electron Transport Complex II / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics*; D006801:Humans; D007834:Lasers; D042282:Microdissection; D008928:Mitochondria / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D004272:DNA, Mitochondrial; D042963:Electron Transport Complex II; D003576:Electron Transport Complex IV