Formation and processing of stalled replication forks--utility of two-dimensional agarose gels.

Journal Article (Journal Article)

Replication forks can be stalled by tightly bound proteins, DNA damage, nucleotide deprivation, or defects in the replication machinery. It is now appreciated that processing of stalled replication forks is critical for completion of DNA replication and maintenance of genome stability. In this chapter, we detail the use of two-dimensional (2D) agarose gels with Southern hybridization for the detection and analysis of blocked replication forks in vivo. This kind of 2D gel electrophoresis has been used extensively for analysis of replication initiation mechanisms for many years, and more recently has become a valuable tool for analysis of fork stalling. Although the method can provide valuable information when forks are stalled in random locations (e.g., after UV damage or nucleotide deprivation), it is even more informative with site-specific fork blockage, for example, blocks caused by tightly bound replication terminator proteins or by drug-stabilized topoisomerase cleavage complexes.

Full Text

Duke Authors

Cited Authors

  • Pohlhaus, JR; Kreuzer, KN

Published Date

  • 2006

Published In

Volume / Issue

  • 409 /

Start / End Page

  • 477 - 493

PubMed ID

  • 16793419

International Standard Serial Number (ISSN)

  • 0076-6879

Digital Object Identifier (DOI)

  • 10.1016/S0076-6879(05)09028-2


  • eng

Conference Location

  • United States