title: Cycling temperature capillary electrophoresis: A quantitative, fast and inexpensive method to detect mutations in mixed populations of human mitochondrial DNA.
abstract: Cycling temperature capillary electrophoresis has been optimised for mutation detection in 76% of the mitochondrial genome. The method was tested on a mixed sample and compared to mutation detection by next generation sequencing. Out of 152 fragments 90 were concordant, 51 discordant and in 11 were semi-concordant. Dilution experiments show that cycling capillary electrophoresis has a detection limit of 1-3%. The detection limit of routine next generation sequencing was in the ranges of 15 to 30%. Cycling temperature capillary electrophoresis detect and accurate quantify mutations at a fraction of the cost and time required to perform a next generation sequencing analysis.
authors.affiliation: École Polytechnique Fédérale de Lausanne, EPFL - FSB - STAP, Station 8, Lausanne, Switzerland. Electronic address: paulo.refinetti@epfl.ch.
authors.forename: Paulo
authors.initials: P
authors.lastname: Refinetti
authors.affiliation: École Polytechnique Fédérale de Lausanne, EPFL - FSB - STAP, Station 8, Lausanne, Switzerland.
authors.forename: Stephan
authors.initials: S
authors.lastname: Morgenthaler
authors.affiliation: Department of Tumor Biology, Norwegian Radiumhospital, Oslo, Norway.
authors.forename: Per O
authors.initials: PO
authors.lastname: Ekstrøm
mesh_terms: D003365:Costs and Cost Analysis; D004272:DNA, Mitochondrial / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000191:economics / Q000379:methods*; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000191:economics; D006801:Humans; D009154:Mutation; D013696:Temperature; D013997:Time Factors
chemical_list: D004272:DNA, Mitochondrial
keywords: Cycling temperature capillary electrophoresis; Heteroplasmy quantification; High throughput; Mitochondrial DNA; Quantitative biology