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Dispersion coefficients of a protein and DNA fragment in polyacrylamide gel electrophoresis as a function of parameters defining the effective gel pore size and particle size
Author(s) -
Chang HuanTsung,
Chrambach Andreas
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501601150
Subject(s) - dispersion (optics) , gel electrophoresis of nucleic acids , particle size , gel electrophoresis , materials science , polyacrylamide gel electrophoresis , chromatography , dna , fragment (logic) , particle (ecology) , electrophoresis , color marker , gel electrophoresis of proteins , molecular weight size marker , chemistry , analytical chemistry (journal) , biology , optics , biochemistry , physics , enzyme , ecology , programming language , computer science
The dispersion coefficient, D ′, of the representative homogeneous protein, conalbumin, decreases linearly as the polyacrylamide concentration increases from 4 to 14%T (2%C), and varies in a biphasic fashion as %C (Bis) is increased from 2 to 20%, with a broad peak between 5 and 15%C. D ′ increases linearly with the concentration of the initiator, potassium persulfate, in the range of 0.01–0.15%. D ′ remains constant when the field strength is varied from 5 to 15 V/cm. A DNA fragment (1857 bp) exhibits a constant D ′ in 4–6% polyacrylamide (2%C) at a field strength of 1 V/cm, and a linearly increasing D ′ at 5 V/cm, in analogy to its previously observed behavior in agarose gels. In solutions of uncrosslinked polyacrylamide, the decrease of the D ′ of conalbumin with polymer concentration is not significantly different from that in 2% N,N ′‐methylenebisacrylamide‐crosslinked gels in the range of 4–14%T, while the decrease of mobility with polyacrylamide concentration is much steeper in 2% crosslinked compared to uncrosslinked polymer. Finally, –∂ (log D ′)/∂ T was found to be proportional to the retardation coefficient, K R (= –∂ (log μ)/∂ T ), in polyacrylamide gels. The ratio of –∂ (log D ′)/∂ T over K R increases with field strength in the range of 5–15 V/cm.