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Development of a Real‐time PCR Assay to Quantify Mitochondrial DNA Deletions Relative to a Housekeeping Gene
Author(s) -
Pitarresi Jason,
Pogozelski Wendy
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.660.6
Subject(s) - housekeeping gene , mitochondrial dna , dna , biology , gene , real time polymerase chain reaction , microbiology and biotechnology , base pair , genetics , nuclear dna , genome , nuclear gene , gene expression
Mitochondria contain DNA in the form of a double‐stranded circle. This DNA (mtDNA) is often found to contain large‐scale deletions of several hundred to several thousand base pairs. These deletions correlate with a variety of disorders in oxidative phosphorylation. Using a real‐time PCR assay based on an external cloned mtDNA quantification standard, we observed that a particular deletion increases in cells treated with gamma rays. However, the data have not always been consistent. We are examining the use of a parallel approach that involves real‐time PCR but measures the deletions and the total mtDNA relative to a nuclear DNA gene. This research was sponsored by the Camille and Henry Dreyfus Foundation, the Geneseo Foundation and the Geneseo Student Association.