Mitochondrial AnalysisOverviewMutations of the mitochondrial genome are the cause of various disorders. Many of them involve multiple organ systems and present with prominent neurological and/or myopathic features. Development of diabetes type II is also frequently observed. Mitochondrial DNA mutations have further been associated with cancer and aging. Denaturing High Performance Liquid Chromatography (DHPLC) in combination with the WAVE® System enables the rapid detection of known and unknown mutations with highest accuracy and specificity by a fully automated procedure involving heteroduplex analysis. A further advantage of the technology is its ability to detect small proportions of mutant alleles within a large excess of wild type DNA. This high level of sensitivity, which is particularly relevant in revealing heteroplasmic mtDNA mutations, has explicitly been documented in the context of mtDNA mutation discovery. Van den Bosch et al. have shown that the WAVE System allows the detection of low levels of heteroplasmic mutations (< 1% heteroplasmy for certain mutations).1 Liu et al. consequently concluded that "DHPLC is a powerful, rapid and sensitive mutation screening method for mtDNA. Proofreading DNA polymerase is more suitable for DHPLC analysis than Taq polymerase."2 References- Van den Bosch BJC, de Coo RFM, Scholte HR, Nijland JG, van den Bogaard R, de Visser M, de Die-Smulders CEM, Smeets HJM. (2000) Mutation analysis of the entire mitochondrial genome using denaturing high performance liquid chromatography. Nucleic Acids Research, Vol. 28, e89. Abstract Library Entry
- Liu M.R., Pan K.F., Li Z.F., Wang Y., Deng D.J., Zhang L., Lu Y.Y. (2002) Rapid screening mitochondrial DNA mutation by using denaturing high-performance liquid chromatography. World J. Gastroenterol., 8, 426-430. Abstract Library Entry
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