The two-dimensional agarose gel electrophoresis (2D gel) is a powerful method used to detect and analyze rare DNA replication and recombination intermediates within a genomic DNA preparation. The 2D gel method has been extensively applied to the budding yeast Saccharomyces cerevisiae due to its small and well-characterized genome to analyze replication fork dynamics at single DNA loci under both physiological and pathological conditions. Here we describe procedures to extract genomic DNA from in vivo UV-psoralen cross-linked yeast cells, to separate branched DNA replication and recombination intermediates by neutral–neutral 2D gel method and to visualize 2D gel structures by Southern Blot.
CITATION STYLE
Zardoni, L., Nardini, E., & Liberi, G. (2020). 2D Gel Electrophoresis to Detect DNA Replication and Recombination Intermediates in Budding Yeast. In Methods in Molecular Biology (Vol. 2119, pp. 43–59). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0323-9_4
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