Title
Trapping of megabase-sized DNA molecules during agarose gel electrophoresis
Date Issued
19 January 1999
Access level
open access
Resource Type
journal article
Author(s)
University of California,
Abstract
Megabase DNA molecules become trapped in agarose gels during electrophoresis if the electric field exceeds a few volts per cm. Fluorescence microscopy reveals that these molecules invariably arrest in U- shaped conformations. The field-vs.-size dependence for trapping indicates that a critical molecular tension is required for trapping. The size of unligated λ-ladders, sheared during gel electrophoresis at a given field, coincides with the size of molecules trapped at that field, suggesting that both processes occur through nick melting near the vertex of the U-shape. Consistently, molecules nicked by exposure to UV radiation trap more readily than unexposed ones. The critical trapping tension at the vertex is estimated to be 15 pN, a force sufficient to melt nicks bent around gel fibers, and, according to our model, trap a molecule. Strategies to reduce molecular tension and avoid trapping are discussed.
Start page
453
End page
458
Volume
96
Issue
2
Language
English
OCDE Knowledge area
Bioquímica, Biología molecular
Genética, Herencia
Scopus EID
2-s2.0-0033582267
PubMed ID
Source
Proceedings of the National Academy of Sciences of the United States of America
ISSN of the container
00278424
Sponsor(s)
National Institute of General Medical Sciences R37GM032543 NIGMS
Sources of information:
Directorio de Producción Científica
Scopus