title: Detection of DNA mutations associated with mitochondrial diseases by Agilent 2100 bioanalyzer.
abstract: BACKGROUND
  Molecular analysis of mitochondrial DNA (mtDNA) has provided a final diagnosis for many of the mitochondrial diseases. We evaluated the Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA) to determine whether the system could replace the conventional restriction fragment length polymorphism (RFLP) analysis by the agarose gel electrophoresis for the detection of the mtDNA mutation.
  
  
  METHODS
  Three members of a family with MELAS syndrome and four members of a family with MERRF syndrome were recruited for this study. After PCR and restriction enzyme digestion, DNA fragments were separated on the Agilent 2100 bioanalyzer in conjunction with the DNA 500 and DNA 1000 Labchip kits and by electrophoresis on precast 3% agarose gels.
  
  
  RESULTS
  The data generated by the DNA 500 and DNA 1000 assays using the Agilent 2100 bioanalyzer showed a lower percentage error and a better reproducibility as compared to those obtained by the conventional method.
  
  
  CONCLUSIONS
  Based on the performance of the bioanalyzer, we suggest that this novel Labchip is adequate to replace the current RFLP analysis by the agarose gel electrophoresis for mtDNA mutation detection.
authors.affiliation: Department of Biochemistry and Center for Cellular and Molecular Biology, School of Life Science, National Yang-Ming University, Taipei, Taiwan 112, ROC.
authors.forename: Ching-You
authors.initials: CY
authors.lastname: Lu
authors.forename: Dan-Ju
authors.initials: DJ
authors.lastname: Tso
authors.forename: Tom
authors.initials: T
authors.lastname: Yang
authors.forename: Yuh-Jyh
authors.initials: YJ
authors.lastname: Jong
authors.forename: Yau-Huei
authors.initials: YH
authors.lastname: Wei
mesh_terms: D000293:Adolescent; D000328:Adult; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000235:genetics* / Q000302:isolation & purification; D004586:Electrophoresis / Q000295:instrumentation*; D005260:Female; D005838:Genotype; D006801:Humans; D017241:MELAS Syndrome / Q000235:genetics; D017243:MERRF Syndrome / Q000235:genetics; D008297:Male; D008838:Microcomputers; D028361:Mitochondrial Diseases / Q000235:genetics*; D018908:Muscle Weakness / Q000235:genetics / Q000473:pathology; D010375:Pedigree; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D004272:DNA, Mitochondrial