title: Recovery of mitochondrial DNA from blood leukocytes using detergent lysis.
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 micrograms 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.
authors.affiliation: Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.
authors.forename: G L
authors.initials: GL
authors.lastname: Lindberg
authors.forename: C M
authors.initials: CM
authors.lastname: Koehler
authors.forename: J E
authors.initials: JE
authors.lastname: Mayfield
authors.forename: A M
authors.initials: AM
authors.lastname: Myers
authors.forename: D C
authors.initials: DC
authors.lastname: Beitz
mesh_terms: D000818:Animals; D002417:Cattle; D004272:DNA, Mitochondrial / Q000097:blood / Q000302:isolation & purification*; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D005821:Genetic Techniques; D007962:Leukocytes / Q000737:chemistry*; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000378:metabolism / Q000494:pharmacology; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D004272:DNA, Mitochondrial; D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific