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The sieving of spheres during agarose gel electrophoresis: Quantitation and modeling
Author(s) -
Griess Gary A.,
Moreno Elena T.,
Easom Richard A.,
Serwer Philip
Publication year - 1989
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.360280811
Subject(s) - agarose , chemistry , radius , electrophoresis , chromatography , analytical chemistry (journal) , hydrodynamic radius , spheres , polymer , physics , organic chemistry , computer security , computer science , copolymer , astronomy
By use of agarose gel electrophoresis, the sieving of spherical particles in agarose gels has been quantitated and modeled for spheres with a radius ( R ) between 13.3 and 149 mm. For quantitation, the electrophoretic mobility has been determined as a function of agarose percentage ( A ). Because a previously used model of sieving [D. Rodbard and A. Chrambach (1970) Proc. Natl. Acad. Sci. USA 65 , 970–977] was found incompatible with some of these data, alternative models have been tested. By use of an underivatized agarose, two models, both based on the assumption of a single effective pore radius ( P E ) for each A , were found to yield P E values that were independent of R and that were in agreement with values of P E obtained independently ( P E = 118 nm × A −0.74 ): sieving by altered hydrodynamics in a cylindrical tube of radius, P E , and sieving by steric exclusion from a circular hole of radius, P E . The same analysis applied to a 6.5% hydroxyethylated commercial agarose yileded a steeper P E vs A plot and also agreement of the above tow models with the data. The P E vs A plot was significantly altered by both further hydroxyethylation and factors that cause variation in the electro‐osmosis found in commercial agarose.