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FORK-seq: replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing.
Genome Biol, 21(1):125.
Genome replication mapping methods profile cell populations, masking cell-to-cellheterogeneity. Here, we describe FORK-seq, a nanopore sequencing method to mapreplication of single DNA molecules at 200-nucleotide resolution. By quantifyingBrdU incorporation along pulse-chased replication intermediates from Saccharomycescerevisiae, we orient 58,651 replication tracks reproducing population-basedreplication directionality profiles and map 4964 and 4485 individual initiation andtermination events, respectively. Although most events cluster at known origins andfork merging zones, 9% and 18% of initiation and termination events, respectively,occur at many locations previously missed. Thus, FORK-seq reveals the full extent ofcell-to-cell heterogeneity in DNA replication.
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