Electric and hydrodynamic stretching of DNA-polymer conjugates in free-solution electrophoresis

• Published in: The Journal of chemical physics Vol. 126; no. 17; p. 175104
• Main Authors: Nedelcu, S, Meagher, R J, Barron, A E, Slater, G W
• Format: Journal Article
• Language: English
• Published: United States 07.05.2007
• Subjects: DNA, Single-Stranded - chemistry; Electromagnetic Fields; Electrophoresis, Polyacrylamide Gel - methods; Models, Theoretical; Polymers - chemistry
• ISSN: 0021-9606
• DOI: 10.1063/1.2730799

The conjugation of an uncharged polymer to DNA fragments makes it possible to separate DNA by free-solution electrophoresis. This end-labeled free-solution electrophoresis method has been shown to successfully separate ssDNA with single monomer resolution up to about 110 bases. It is the aim of this paper to investigate in more detail the coupled hydrodynamic and electrophoretic deformation of the ssDNA-label conjugate at fields below 400 V/cm. Our model is an extension of the theoretical approach originally developed by Stigter and Bustamante [Biophys. J. 75, 1197 (1998)] to investigate the problems of a tethered chain stretching in a hydrodynamic flow and of the electrophoretic stretch of a tethered polyelectrolyte. These two separate models are now used together since the charged DNA is "tethered" to the uncharged polymer (and vice versa), and the resulting self-consistent model is used to predict the deformation and the electrophoretic velocity for the hybrid molecule. Our theoretical and experimental results are in good qualitative agreement.