Direct quantification of mitochondrial DNA and its 4.9-kb common deletion without DNA purification
| Autor: | A. Dib, André Peinnequin, C. Demeilliers, C. Mouret, T. Poyot, A. Aubourg |
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| Přispěvatelé: | Centre de recherche biomédicale Bichat-Beaujon (CRB3), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM) |
| Rok vydání: | 2011 |
| Předmět: |
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT]
Mitochondrial DNA Lysis Biophysics Biology DNA Mitochondrial Polymerase Chain Reaction Biochemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Animals Rats Wistar Molecular Biology ComputingMilieux_MISCELLANEOUS Sequence Deletion 030304 developmental biology 0303 health sciences Cell Biology Molecular biology DNA extraction Rats genomic DNA Liver chemistry Quantitative analysis (chemistry) 030217 neurology & neurosurgery DNA |
| Zdroj: | Analytical Biochemistry Analytical Biochemistry, Elsevier Masson, 2011, 409 (2), pp.298-300. ⟨10.1016/j.ab.2010.10.010⟩ |
| ISSN: | 0003-2697 1096-0309 |
| DOI: | 10.1016/j.ab.2010.10.010 |
| Popis: | Quantitative analysis of mitochondrial DNA (mtDNA) and its common deletion (CD) are sensitive and early markers for mitochondrial mutations and suffering. However, the use of purified DNA can lead to quantification errors because of variable DNA extraction yields due to the significant differences in size and structure between genomic DNA (gDNA) and mtDNA. We report a real-time qPCR-based protocol directly on tissue lysate, without DNA extraction. This method, which allows both absolute and relative measure, increases the measuring accuracy of the mtDNA/gDNA ratio and leads to reliable and more reproducible results when measuring the deleted/total mtDNA ratio. |
| Databáze: | OpenAIRE |
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