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Proceedings Paper

Fluorescence microscopy and computer simulation studies of the mechanisms of reorientation of DNA molecules undergoing pulsed-field gel electrophoresis
Author(s): Carlos J. Bustamante; Steven B. Smith; Sergio Gurrieri
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Paper Abstract

The mechanisms of reorientation of individual DNA molecules undergoing Pulsed Field Gel Electrophoresis (PFGE) have been studied using T2 DNA molecules labeled with acridine orange and visualized with a fluorescence microscope. It is shown that molecules undergoing PFGE and conventional electrophoresis often get trapped in hook conformations (narrow U-shapes) that play an important role in determining the mobility of the molecules. It is found that the mechanism of formation of hooks require the previous generation of a kink (in which parts of the molecule double up inside a pore). Computer simulation experiments are presented to clarify the role of hook and kink formation in the size-dependent separation observed in PFGE experiments.

Paper Details

Date Published: 1 August 1990
PDF: 13 pages
Proc. SPIE 1205, Bioimaging and Two-Dimensional Spectroscopy, (1 August 1990); doi: 10.1117/12.17800
Show Author Affiliations
Carlos J. Bustamante, Univ. of New Mexico (United States)
Steven B. Smith, Univ. of New Mexico (United States)
Sergio Gurrieri, Univ. of New Mexico (United States)


Published in SPIE Proceedings Vol. 1205:
Bioimaging and Two-Dimensional Spectroscopy
Louis C. Smith, Editor(s)

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