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Optimization of separation conditions in the size-selective capillary electrophoretic separation of DNA fragments

1996; Wiley; Volume: 8; Issue: 7 Linguagem: Inglês

10.1002/(sici)1520-667x(1996)8

1520-667X

Michael Stebbins, Joe B. Davis, B. K. Clark, Michael J. Sepaniak,

Microfluidic and Bio-sensing Technologies

We report herein the development and initial evaluation of a computer program to determine optimal separation conditions for the size-selective capillary electrophoretic (SSCE) separation of DNA fragments. The program uses the simplex method to optimize for those conditions which are most influential in these types of separations. Since base-pair electrophoretic behavior is largely independent of sequence, an empirical equation can be used to predict DNA fragment mobility in methyl cellulose running buffers. This allows column length and methyl cellulose concentration to be adjusted for optimal separation performance. The program generates a chromatographic response function which serves as a measure of the success of the separation. Reasonable correlation between computational and experimental work is demonstrated, as are the influences of adjustable parameters on the quality of the separation and the function of the computational methods. Also presented is the ability of the program to function for samples that were not used in the initial data set. © 1996 John Wiley & Sons, Inc. J Micro Sep 8: 485–494, 1996