A new chapter for ERAD?

Cunliffe, Lesley

doi:10.1038/nrm1776

Nat Rev Mol Cell Biol

2005

DOI: 10.1038/nrm1776

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A new chapter for ERAD?

Lesley Cunliffe¹

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2007

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Quantitation of DNA Copy Number in Individual Mitochondrial Particles by Capillary Electrophoresis

2007

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Here, we present a direct method for determining mitochondrial DNA (mtDNA) copy numbers in individual mitochondrial particles, isolated from cultured cells, by means of capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection. We demonstrate that this method can detect a single molecule of PicoGreen-stained mtDNA in intact DsRed2-labeled mitochondrial particles isolated from human osteosarcoma 143B cells. This ultimate limit of mtDNA detection made it possible to confirm that an individual mitochondrial nucleoid, the genetic unit of mitochondrial inheritance, can contain a single copy of mtDNA. The validation of this approach was achieved via monitoring chemically induced mtDNA depletion and comparing the CE-LIF results to those obtained by quantitative microscopy imaging and multiplex real-time PCR analysis. Owing to its sensitivity, the CE-LIF method may become a powerful tool for investigating the copy number and organization of mtDNA in mitochondrial disease and aging, and in molecular biology techniques requiring manipulation and quantitation of DNA molecules such as plasmids.

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Quantitation of DNA Copy Number in Individual Mitochondrial Particles by Capillary Electrophoresis

2007

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A new chapter for ERAD?

Cited by 1 publication

References 3 publications

Quantitation of DNA Copy Number in Individual Mitochondrial Particles by Capillary Electrophoresis

Quantitation of DNA Copy Number in Individual Mitochondrial Particles by Capillary Electrophoresis

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