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Transverse imaging and simulation of dsDNA electrophoresis in microfabricated glass channels
Author(s) -
Zeitoun Ramsey I.,
Chen Zheng,
Burns Mark A.
Publication year - 2008
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.200800204
Subject(s) - electrophoresis , gel electrophoresis of nucleic acids , confocal , transverse plane , dna , confocal microscopy , gel electrophoresis , chemistry , distribution (mathematics) , biophysics , molecule , microscopy , materials science , analytical chemistry (journal) , optics , chromatography , physics , biology , anatomy , biochemistry , mathematical analysis , mathematics , organic chemistry
We have observed the non‐uniform distribution of DNA molecules during PAGE in a microfabricated system. Confocal laser scanning microscopy was used to visualize fluorescently labeled DNA during electrophoretic migration. The distribution of double‐stranded DNA larger than 100 bp is observed to transition from a center‐biased motion on the transverse plane 1 cm downstream from injection to an edge‐biased motion 2 cm downstream. Although this distribution increased with increasing dsDNA size in a cross‐linked gel, no similar distribution was found with the same dsDNA molecules in a linear polyacrylamide solution (6%). Simulations of DNA distribution in gels suggest that DNA distribution non‐uniformities may be caused by biased electrophoretic migration resulting from motion in an inhomogeneous gel system.