title: Quantitation of Mitochondrial DNA Deletions Via Restriction Digestion/Long-Range Single-Molecule PCR.
abstract: Quantification of deletions in mtDNA is a long-standing problem in mutational analysis. We describe here an approach that combines the power of single-molecule PCR of the entire mitochondrial genome with the enrichment of the deletions by restriction digestion. This approach is indispensable if information about wide range of deletion types in a sample is critical, such as in studies concerning distribution of deletion breakpoints (as opposed to approaches where fraction of a single deletion or a limited set of deletions is used as a proxy for total deletion load). Because deletions in a sample are quantified almost exhaustively, the other important application of this approach involves studies where only small amounts of tissue, such as biopsies, are available.
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Yevgenya
authors.initials: Y
authors.lastname: Kraytsberg
authors.affiliation: College of Biology, Hunan University, Changsha, People's Republic of China.
authors.forename: Xinhong
authors.initials: X
authors.lastname: Guo
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Saisai
authors.initials: S
authors.lastname: Tao
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Kuznetsov
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: MacLean
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: McLean
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Ehrlich
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Evan
authors.initials: E
authors.lastname: Feldman
authors.affiliation: Beth Israel Deaconess Medical Center, Boston, MA, USA.
authors.forename: Igor
authors.initials: I
authors.lastname: Dombrovsky
authors.affiliation: Heart Centre, The Affiliated Hospital, Hangzhou Normal University, Hangzhou, People's Republic of China.
authors.forename: Deye
authors.initials: D
authors.lastname: Yang
authors.affiliation: Bouve College of Health Sciences, Northeastern University, Boston, MA, USA.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Cloutier
authors.affiliation: Bouve College of Health Sciences, Northeastern University, Boston, MA, USA.
authors.forename: Carmen
authors.initials: C
authors.lastname: Castaneda-Sceppa
authors.affiliation: Department of Biology, Northeastern University, 300 Huntington Avenue, Boston, MA, 02115, USA. kkhrapko@gmail.com.
authors.forename: Konstantin
authors.initials: K
authors.lastname: Khrapko
mesh_terms: D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D004252:DNA Mutational Analysis / Q000379:methods*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics*; D054629:Genome, Mitochondrial / Q000235:genetics*; D006801:Humans; D028361:Mitochondrial Diseases / Q000235:genetics; D009132:Muscles / Q000166:cytology; D009206:Myocardium / Q000166:cytology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018384:Oxidative Stress / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D017382:Reactive Oxygen Species / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics; D013378:Substantia Nigra / Q000166:cytology
chemical_list: D004272:DNA, Mitochondrial; D017382:Reactive Oxygen Species; D004262:DNA Restriction Enzymes
keywords: Deletions; Exercise; Mitochondrial DNA; Mutation; PCR; Restriction digestion