Abstract
Mitochondrial DNA (mtDNA) was isolated from leukocytes contained in whole blood of cattle. Leukocyte membranes except the nuclear envelope were solubilized in a buffer that contained 1% Triton X-100. After sedimentation of cell nuclei, mtDNA was purified from the cell lysate by organic solvent extraction and ethanol precipitation. Approximately 5 µg of mtDNA was recovered from 400 ml of whole blood, a quantity sufficient for routine DNA cloning procedures or for detailed restriction mapping studies. mtDNA isolated with this method is a suitable substrate for several DNA-modifying enzymes. Thus, preparation of mtDNA from blood by detergent lysis provides a noninvasive alternative to tissue biopsy for characterization of mitochondrial genotypes in studies of evolutionary genetics and population dynamics.
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References
Anderson, S., de Bruijn, M. H. L., Coulson, A. R., Eperon, I. C., Sanger, F., and Young, I. G. (1982). The complete sequence of bovine mitochondrial DNA: Conserved features of the mammalian mitochondrial genome.J. Mol. Biol. 156683.
Avise, J. C., Arnold, J., Bell, R. M., Bermingham, E., Lamb, T., Neigel, J. E., Reeb, C. A., and Saunders, N. C. (1987). Intraspecific phylogeography: The mitochondrial DNA bridge between population genetics and systematics.Annu. Rev. Ecol. Syst. 18489.
Birnboim, H., and Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA.Nucl. Acids Res. 71513.
Brown, W. M., and Vinograd, J. (1974). Restriction endonuclease cleavage maps of animal mitochondrial DNAs.Proc. Natl. Acad. Sci. USA 714617.
Hixson, J. E., Wong, T. T., and Clayton, D. A. (1986). Both the conserved stem-loop and divergent 5′-flanking sequences are required for initiation at the human mitochondrial origin of light-strand DNA replication.J. Biol. Chem. 2612384.
Holt, I. J., Harding, A. E., and Morgan-Hughes, J. A. (1988). Deletions of muscle mitochondrial DNA in patients with mitochondrial myopathies.Nature 331717.
Hymer, W. C., and Kuff, E. L. (1964). Isolation of nuclei from mammalian tissues through the use of Triton X-100.J. Histochem. Cytochem. 12359.
Koehler, C. M., Lindberg, G. L., Brown, D. R., Beitz, D. C., Mayfield, J. E., Freeman, A. E., and Myers, A. M. (1991). Replacement of bovine mitochondrial DNA by a sequence variant within one generation.Genetics 121247.
Lindberg, G. L. (1989).Sequence Heterogeneity of Bovine Mitochondrial DNA Ph.D. dissertation, Parks Library, Iowa State University, Ames.
Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Manual Cold Spring Laboratory, Cold Spring Harbor, N.Y.
Roos, D., and Loos, J. A. (1970). Changes in carbohydrate metabolism of mitogenically stimulated human peripheral lymphocytes. I. Stimulation by phytohemagglutinin.Biochim. Biophys. Acta 222565.
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This work was supported in part by the Iowa State University Biotechnology Council and by the Holstein Association.
Journal Paper No. J-13683 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 2736.
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Lindberg, G.L., Koehler, C.M., Mayfield, J.E. et al. Recovery of mitochondrial DNA from blood leukocytes using detergent lysis. Biochem Genet 30, 27–33 (1992). https://doi.org/10.1007/BF00554425
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DOI: https://doi.org/10.1007/BF00554425