Using ΦX174 DNA as an exogenous reference for measuring mitochondrial DNA copy number
作者： Meagan B. Myers , Roberta A. Mittelstaedt , Robert H. Heflich
DOI: 10.2144/000113222
关键词: DNA 、 Mitochondrial DNA 、 Gene dosage 、 Real-time polymerase chain reaction 、 Polymerase chain reaction 、 DNA extraction 、 Nuclear DNA 、 Biology 、 Genetics 、 Ploidy
摘要: Quantitative real-time PCR has become a popular method to analyze and quantify changes in the copy number of mitochondrial DNA (mtDNA), nuclear (nDNA) is often used as an endogenous reference for mtDNA abundance. In our experience, using nDNA problematic, due differences extraction efficiency variation ploidy experimental samples. Here, we report that ratio varies repeated extractions but ΦX174 DNA, added before extraction, extracted with similar mtDNA, making it suitable alternative quantifying number.
future-science.com
unirioja.es
biotechniques.com 参考文章(13)
Franck Chiappini, Elina Teicher, Raphaël Saffroy, Patrick Pham, Bruno Falissard, Alain Barrier, Stephan Chevalier, Brigitte Debuire, Daniel Vittecoq, Antoinette Lemoine, Prospective evaluation of blood concentration of mitochondrial DNA as a marker of toxicity in 157 consecutively recruited untreated or HAART-treated HIV-positive patients Laboratory Investigation. ,vol. 84, pp. 908- 914 ,(2004) , 10.1038/LABINVEST.3700113
Kenneth J. Livak, Thomas D. Schmittgen, ANALYSIS OF RELATIVE GENE EXPRESSION DATA USING REAL-TIME QUANTITATIVE PCR AND THE 2(-DELTA DELTA C(T)) METHOD Methods. ,vol. 25, pp. 402- 408 ,(2001) , 10.1006/METH.2001.1262
Thomas D. Schmittgen, Brian A. Zakrajsek, Alan G. Mills, Vladimir Gorn, Michael J. Singer, Michael W. Reed, Quantitative Reverse Transcription–Polymerase Chain Reaction to Study mRNA Decay: Comparison of Endpoint and Real-Time Methods☆ Analytical Biochemistry. ,vol. 285, pp. 194- 204 ,(2000) , 10.1006/ABIO.2000.4753
E.A. Schon, C.T. Moraes, E.A. Schon, M. Dalakas, E. Arnaudo, S. Shanske, S. DiMauro, Depletion of muscle mitochondrial DNA in AIDS patients with zidovudine-induced myopathy The Lancet. ,vol. 337, pp. 508- 510 ,(1991) , 10.1016/0140-6736(91)91294-5
F. J. Miller, Precise determination of mitochondrial DNA copy number in human skeletal and cardiac muscle by a PCR‐based assay: lack of change of copy number with age Nucleic Acids Research. ,vol. 31, ,(2003) , 10.1093/NAR/GNG060
Margaret Keighren, John D. West, Analysis of cell ploidy in histological sections of mouse tissues by DNA-DNA in situ hybridization with digoxigenin-labelled probes. Histochemical Journal. ,vol. 25, pp. 30- 44 ,(1993) , 10.1007/BF00161042
Eric A. Schon, June R. Aprille, Salvatore DiMauro, Carlos T. Moraes, Eduardo Bonilla, Sara Shanske, Francesca Andreetta, Hans Jürgen Tritschler, mtDNA depletion with variable tissue expression: a novel genetic abnormality in mitochondrial diseases. American Journal of Human Genetics. ,vol. 48, pp. 492- 501 ,(1991)
Sanjeev Gupta, Hepatic polyploidy and liver growth control Seminars in Cancer Biology. ,vol. 10, pp. 161- 171 ,(2000) , 10.1006/SCBI.2000.0317
Hélène CF Côté, Zabrina L Brumme, Kevin JP Craib, Christopher S Alexander, Brian Wynhoven, Lillian Ting, Hubert Wong, Marianne Harris, P Richard Harrigan, Michael V O'Shaughnessy, Julio SG Montaner, None, Changes in Mitochondrial DNA as a Marker of Nucleoside Toxicity in HIV-Infected Patients The New England Journal of Medicine. ,vol. 346, pp. 811- 820 ,(2002) , 10.1056/NEJMOA012035
William Lewis, Brian J. Day, William C. Copeland, Mitochondrial toxicity of NRTI antiviral drugs: an integrated cellular perspective. Nature Reviews Drug Discovery. ,vol. 2, pp. 812- 822 ,(2003) , 10.1038/NRD1201