Sieving of double-stranded DNA during agarose gel electrophoresis

Abstract

Agarose gel electrophoresis is used to fractionate linear, double-stranded DNA by its length. Sieving of the gel is the cause of this fractionation and has been investigated by developing theoretical models and by quantifying sieving observed during electrophoresis. Here are reviewed the following aspects of the fractionation of linear, double-stranded DNA by agarose gel electrophoresis: (1) the basic observations that qualitatively characterize these fractionations, (2) evidence for the deformation of DNA's random coil, (3) quantitative analysis of the relationship of observed electrophoretic mobility to the DNA's length, (4) theoretical models that have been developed to explain data presented in Sections 1-3, (5) observations not yet quantitatively explained by models, and (6) some aspects of the use of a variable electrical field (pulsed-field gel electrophoresis) to improve separations.