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Mapping DNA replication with nanopore sequencing


We have harnessed nanopore sequencing to study DNA replication genome-wide at the single-molecule level. Using in vitro prepared DNA substrates, we characterized the effect of bromodeoxyuridine (BrdU) substitution for thymidine on the MinION nanopore electrical signal. Using a neural-network basecaller trained on yeast DNA containing either BrdU or thymidine, we identified BrdU-labelled tracts in yeast cells synchronously entering S phase in the presence of hydroxyurea and BrdU. As expected, the BrdU-labelled tracts coincided with previously identified early-firing, but not late-firing, replication origins. These results open the way to high-throughput, high-resolution, single-molecule analysis of DNA replication in many experimental systems.

Authors: Magali Hennion, Jean-Michel Arbona, Corinne Cruaud, Florence Proux, Benoît Le Tallec, Elizaveta Novikova, Stéfan Engelen, Arnaud Lemainque, Benjamin Audit, Olivier Hyrien

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