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.
CITATION STYLE
Hennion, M., Arbona, J. M., Lacroix, L., Cruaud, C., Theulot, B., Tallec, B. L., … Hyrien, O. (2020). FORK-seq: Replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing. Genome Biology, 21(1). https://doi.org/10.1186/s13059-020-02013-3
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