Multiplex Capillary Denaturing High-Performance Liquid Chromatography with Laser-Induced Fluorescence Detection. (June 2001)

• Record Type: Journal Article
• Authors: Xiao, W.; Stern, D.; Jain, M.; Huber, C.G.; Oefner, P.J.
• Is Part Of: Biotechniques. Volume 30:Number 6(2001)
• Page Start: 1332
• Page End: 1338
• Publication Date: 2001-06
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/01306rr01
• Languages: English
• ISSNs: 0736-6205

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 throughput, 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.