Single-cell A3243G mitochondrial DNA mutation load assays for segregation analysis

Sammanfattning

Segregation of mitochondrial DNA (mtDNA) is an important underlying pathogenic factor in mtDNA mutation accumulation in mitochondrial diseases and aging, but the molecular mechanisms of mtDNA segregation are elusive. Lack of high-throughput single-cell mutation load assays lies at the root of the paucity of studies in which, at the single-cell level, mitotic mtDNA segregation patterns have been analyzed. Here we describe development of a novel fluorescence-based, non-gel PCR restriction fragment length polymorphism method for single-cell A3243G mtDNA mutation load measurement. Results correlated very well with a quantitative in situ Padlock/rolling circle amplification-based genotyping method. In view of the throughput and accuracy of both methods for single-cell A3243G mtDNA mutation load determination, we conclude that they are well suited for segregation analysis

Nyckelord

Cell Separation; Cells; Cultured; DNA; Mitochondrial/*genetics; Genotype; Humans; Image Processing; Computer-Assisted; Microscopy; Fluorescence; Mutation; Polymerase Chain Reaction/methods; Polymorphism; Restriction Fragment Length; Transition Temperature

Publicerad i

Journal of Histochemistry and Cytochemistry
2007, volym: 55, nummer: 11, sidor: 1159-1166

SLU författare

• Det finns ingen information

Publikationens identifierare

• DOI: https://doi.org/10.1369/jhc.7a7282.2007

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/16156