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Mobility surfaces for field‐inversion gel electrophoresis of linear DNA
Author(s) -
Crater Glenn D.,
Gregg Michael R.,
Holzwarth G.
Publication year - 1989
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.1150100507
Subject(s) - agarose , electrophoresis , agarose gel electrophoresis , physics , gel electrophoresis , analytical chemistry (journal) , field (mathematics) , pulse (music) , dna , chemistry , materials science , mathematics , optics , chromatography , biochemistry , detector , pure mathematics
The mobility of linear DNA during field‐inversion gel electrophoresis was measured as a function of molecular weight M r , pulse time t , and field strength E. Values of M r between 48.5 and 194 kilobase pairs (kb), E from 5 to 14 V/cm and pulse times of 0.3 to 12 s were used. The data are presented as three‐dimensional surfaces of mobility: E:t for fixed M r or graphs of mobility: M r : t for fixed E. The surfaces are not smoothly increasing functions of E , M r , or t but instead show a valley with minimum mobility and a steep rise in mobility as t increases. For a field of 10 V/cm, 1 % agarose gels, and 3:1 ratio of forward: back pulse time, the forward switching time t * at which the mobility changes most rapidly is given by t * =(0.034 ± 0.003) M r for M r in kb and t * in seconds. The data and equations delineate the best conditions to achieve a particular separation.