Fluorescence microscopy and computer simulation studies of the mechanisms of reorientation of DNA molecules undergoing pulsed-field gel electrophoresis

Carlos J. Bustamante; Steven B. Smith; Sergio Gurrieri

doi:10.1117/12.17800

1 August 1990 Fluorescence microscopy and computer simulation studies of the mechanisms of reorientation of DNA molecules undergoing pulsed-field gel electrophoresis

Carlos J. Bustamante, Steven B. Smith, Sergio Gurrieri

Author Affiliations +

Proceedings Volume 1205, Bioimaging and Two-Dimensional Spectroscopy; (1990) https://doi.org/10.1117/12.17800
Event: OE/LASE '90, 1990, Los Angeles, CA, United States

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.

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Carlos J. Bustamante, Steven B. Smith, and Sergio Gurrieri "Fluorescence microscopy and computer simulation studies of the mechanisms of reorientation of DNA molecules undergoing pulsed-field gel electrophoresis", Proc. SPIE 1205, Bioimaging and Two-Dimensional Spectroscopy, (1 August 1990); https://doi.org/10.1117/12.17800

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