RT Journal
A1 Premstaller, Andreas
A1 Xiao, Wenzhong
A1 Oberacher, Herbert
A1 O'Keefe, Matthew
A1 Stern, David
A1 Willis, Thomas
A1 Huber, Christian G.
A1 Oefner, Peter J.
T1 Temperature-Modulated Array High-Performance Liquid Chromatography
JF Genome Research 
JO Genome Research 
YR 2001 
FD November 01 
VO 11 
IS 11 
SP 1944 
OP 1951 
DO 10.1101/gr.200401 
UL http://genome.cshlp.org/content/11/11/1944.abstract 
AB Using novel monolithic poly(styrene-divinylbenzene) capillary columns with an internal diameter of 0.2 mm, we demonstrate for the first time the feasibility of constructing high-performance liquid chromatography arrays for the detection of mutations by heteroduplex analysis under partially denaturing conditions. In one embodiment, such an array can be used to analyze one sample simultaneously at different temperatures to maximize the detection of mutations in DNA fragments containing multiple discrete melting domains. Alternatively, one may inject different samples onto columns kept at the same effective temperature. Further improvements in throughput can be obtained by means of laser-induced fluorescence detection and the differential labeling of samples with up to four different fluorophores. Major advantages of monolithic capillary high-performance liquid chromatographic arrays over their capillary electrophoretic analogs are the chemical inertness of the poly(styrene-divinylbenzene) stationary phase, the physical robustness of the column bed due to its covalent linkage to the inner surface of the fused silica capillary, and the feasibility to modify the stationary phase thereby allowing the separation of compounds not only on the principle of size exclusion, but also adsorption, distribution, and ion exchange. Analyses times are on the order of a few minutes and turnaround time is extremely short as there is no need for the replenishment of the separation matrix between runs.