title: Directionality of replication fork movement determined by two-dimensional native-native DNA agarose gel electrophoresis.
abstract: The analysis of replication intermediates by the neutral-neutral two-dimensional agarose gel technique allows determining the chromosomal positions where DNA replication initiates, whether replication forks pause or stall at specific sites, or whether two DNA molecules undergo DNA recombination events. This technique does not, however, immediately tell in which direction replication forks migrate through the DNA region under investigation. Here, we describe the procedure to determine the direction of replication fork progression by carrying out a restriction enzyme digest of DNA imbedded in agarose after the completion of the first dimension of a 2D gel.
authors.affiliation: Department of Cell Biology and Molecular Medicine, University of Medicine and Dentistry of New Jersey, Newark, NJ, USA.
authors.forename: Andreas S
authors.initials: AS
authors.lastname: Ivessa
mesh_terms: D004247:DNA / Q000737:chemistry*; D004261:DNA Replication / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D018741:Replication Origin; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D004247:DNA; D004262:DNA Restriction Enzymes