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Updated: Dec 29, 2025
Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion
Published on: September 13, 2024
2D Gel Electrophoresis to Detect DNA Replication and Recombination Intermediates in Budding Yeast
Luca Zardoni1,2, Eleonora Nardini1, Giordano Liberi3,4
1Istituto di Genetica Molecolare, CNR, Pavia, Italy.
Abstract:
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.
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