Multiplex capillary denaturing high-performance liquid chromatography with laser-induced fluorescence detection. |
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Authors: | W Xiao D Stern M Jain C G Huber P J Oefner |
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Affiliation: | Stanford University, Palo Alto, CA, Affymetrix, Santa Clara, CA, USA. |
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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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