Systematic study of field and concentration effects in capillary electrophoresis of DNA in polymer solutions.

A systematic study of the separation of double-stranded DNA in hydroxypropylcellulose (HPC) with a molecular mass of 10(6) was undertaken, using a variety of concentrations (from 0.1 to 1%) and different electric fields (from 6 to 540 V/cm). The data show that at high polymer concentrations ( > or = 0.4%) and low fields, the separation mechanism is similar to that occurring in gels. The results are in good agreement with theoretical models, and in particular with a recently proposed theory for gels with a pore size smaller than the persistence length of DNA. For more dilute solutions and high fields, however, the separation pattern cannot be explained by existing theories. The existence of an original mechanism was confirmed by the direct observation of the conformation of double-stranded DNA molecules in the polymer solution by fluorescence videomicroscopy. Practical conclusions for the capillary electrophoretic separation of duplex DNA are drawn.

[1]  M. Strege,et al.  Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries. , 1991, Analytical chemistry.

[2]  Daniel Broseta,et al.  Universal Properties of Semi-Dilute Polymer Solutions: A Comparison between Experiments and Theory , 1986 .

[3]  T. Duke,et al.  Electrophoretic mobility of DNA in gels. II. Systematic experimental study in agarose gels , 1994 .

[4]  T. Duke,et al.  Two‐dimensional motion of DNA bands during 120° pulsed‐field gel electrophoresis. I. Effect of molecular weight , 1995 .

[5]  A. G. Ogston,et al.  The spaces in a uniform random suspension of fibres , 1958 .

[6]  D. Soane,et al.  Experimental and theoretical studies of DNA separations by capillary electrophoresis in entangled polymer solutions , 1991, Biopolymers.

[7]  Norman Davidson,et al.  Electrophoresis of the nucleic acids , 1964 .

[8]  H. Blanch,et al.  Capillary electrophoresis of DNA in uncross-linked polymer solutions. , 1993, Journal of chromatography. A.

[9]  C. P. Bean,et al.  Electrophoretic mobility of λ phage HIND III and HAE III DNA fragments in agarose gels: A detailed study , 1987 .

[10]  T. Duke,et al.  Electrophoretic mobility of DNA in gels. I. New biased reptation theory including fluctuations , 1994 .

[11]  T. Duke,et al.  DNA electrophoresis in polymer solutions: Ogston sieving, reptation and constraint release , 1993, Electrophoresis.

[12]  P. Righetti,et al.  Movement of DNA fragments during capillary zone electrophoresis in liquid polyacrylamide , 1993 .

[13]  S. Edwards,et al.  The Theory of Polymer Dynamics , 1986 .

[14]  Steven B. Smith,et al.  Electrophoretic charge density and persistence length of DNA as measured by fluorescence microscopy , 1990, Biopolymers.