Capillary Electrophoresis—Laser Fluorometry Instrumentation for the Facile Optimization of DNA Separations Using in Situ Size-Selective Gradients and Adjustable Detection Zone

The construction and evaluation of a capillary electrophoresis instrument for the facile optimization of DNA separations is described herein. This instrumentation allows for convenient translation of the detection zone with little loss in detectability. Changing the effective capillary length is shown to be important when complete resolution of specified DNA fragments in a sample, in the minimum time, is desired. Efficiency is found to decrease with column length in accordance with limits imposed by axial diffusion. However, complete resolution of the fragments in a ΦX-174-HeaIII digest is accomplished in 155s at moderate field strengths and an effective capillary length of 15 cm. It is also demonstrated that in situ gradients can be established using this instrumentation. The general chromatographic elution problem is addressed with gradients, resulting in further improvements in separation performance relative to isocratic separations. © 1995, American Chemical Society. All rights reserved.

Duke Scholars

Author Tuan Vo-Dinh Biomedical Engineering