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FORK-seq: replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing

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Hennion, Magali | Arbona, Jean-Michel | Lacroix, Laurent | Cruaud, Corinne | Theulot, Bertrand | Tallec, Benoît, Le | Proux, Florence | Wu, Xia | Novikova, Elizaveta | Engelen, Stefan | Lemainque, Arnaud | Audit, Benjamin | Hyrien, Olivier

Edité par CCSD ; BioMed Central -

International audience. Genome replication mapping methods profile cell populations, masking cell-to-cell heterogeneity. Here, we describe FORK-seq, a nanopore sequencing method to map replication of single DNA molecules at 200-nucleotide resolution. By quantifying BrdU incorporation along pulse-chased replication intermediates from Saccharomyces cerevisiae, we orient 58,651 replication tracks reproducing population-based replication directionality profiles and map 4964 and 4485 individual initiation and termination events, respectively. Although most events cluster at known origins and fork merging zones, 9% and 18% of initiation and termination events, respectively, occur at many locations previously missed. Thus, FORK-seq reveals the full extent of cell-to-cell heterogeneity in DNA replication.

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