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Capillary electrophoresis‐laser induced fluorescence analysis of endogenous damage in mitochondrial and genomic DNA
Author(s) -
Wirtz Michaela,
Schumann Claus A.,
Schellenträger Marc,
Gäb Siegmar,
vom Brocke Jochen,
Podeschwa Michael A. L.,
Altenbach HansJ.,
Oscier David,
Schmitz Oliver J.
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200410397
Subject(s) - mitochondrial dna , dna , biochemistry , genomic dna , microbiology and biotechnology , chemistry , dna damage , capillary electrophoresis , biology , gene
Reactive oxygen molecules are formed in vivo as by‐products of normal aerobic metabolism. All organisms dependent on oxygen are inevitably exposed to these species so that DNA damage can occur in both genomic and mitochondrial DNA (mtDNA). In order to determine endogenous DNA damage we have developed an analytical method that involves the isolation and hydrolysis of genomic DNA or mtDNA, the labeling of modified and unmodified nucleotides and micellar electrokinetic chromatography with laser‐induced fluorescence detection. With this method we have found etheno‐adenine, thymine glycol, uracil, hypoxanthine, and 5‐methylcytosine. These were identified by the addition of internal standards to the genomic or mtDNA. There are a large number of other signals in the electropherograms of mtDNA that we have never found in genomic DNA analysis because they are at lower concentration in the genome. In the DNA of untreated patients with chronic lymphocytic leukemia (CLL), uracil and high levels of etheno‐adenine were found, which can be explained by antioxidant enzyme alterations and oxidative stress in the CLL lymphocytes.