A study on protein and DNA analysis by Capillary electrophoresis

Abstract

The aim of this research was to establish a method for the analysis of several proteins and DNA using capillary electrophoresis. Bovine serum albumin, human serum proteins and some restriction enzymes namely Taq1, EcoR1, Pst1 were analysed by capillary zone electrophoresis in fused silica capillary columns. Entangled solution capillary electrophoresis by applying dynamic coating with the use of a run buffer containing non-cross linked polyacrylamide and capillary gel electrophoresis in polyacrylamide gel-filled columns were performed for the separation of the 491 bp and 488 bp PCR amplified DNA fragments from Turkish cystic fibrosis carriers. The effect of various parameters such as the use of different buffer systems, capillary dimensions, detector wavelengths, polarity modes, applied voltage, sample injection modes and sample loading amounts on the analytical parameters were evaluated. The optimum wavelength for the detection of proteins was 214 nm. Highest voltages possible should be used for the fastest and most efficient separation. The use of a shorter capillary column resulted in faster migration, less zone broadening and narrower peaks. The TBE buffer used for the analysis of restriction enzymes was found to be not suitable while the most appropriate buffer system for the analysis of human serum proteins was Dolnik's buffer. For the analysis of DNA fragments, optimization of the dynamic coating procedure or static coating of the capillary column with linear polyacrylamide should be developed.