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Multiplex capillary denaturing high-performance liquid chromatography with laser-induced fluorescence detection.

W Xiao·D Stern·M Jain·C G Huber·P J Oefner
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Abstract

Denaturing high-performance liquid chromatography (DHPLC) is a sensitive, robust, and operationally inexpensive method for the detection of single-base substitutions and small deletions and insertions. To increase sample throughout, we have developed a multiplexing strategy using fluorophores to distinguish different PCR products. The system combines recent advances in the synthesis of monolithic poly(styrene-divinylbenzene) capillary columns with four-color confocal argon ion laser-induced fluorescence detection. Depending on the change in retention caused by the fluorophores, adjustments in the analysis temperature may be required to ensure the maximum mutation detection sensitivity.

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ArgonBase Pair MismatchChromatography, High Pressure LiquidDNA Mutational AnalysisElectrophoresis, CapillaryEquipment DesignFluorescent DyesFluorometryHeteroduplex AnalysisHumansLasersMaleNucleic Acid DenaturationNucleic Acid HeteroduplexesPolymerase Chain ReactionPolymorphism, GeneticTemperatureY ChromosomeJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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W Xiao, D Stern, M Jain, C G Huber, P J Oefner. 2001. Multiplex capillary denaturing high-performance liquid chromatography with laser-induced fluorescence detection.. https://doi.org/10.2144/01306rr01

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