FORK-seq: Single-Molecule Profiling of DNA Replication
Résumé
Most 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 using a nanopore current interpretation tool allowing the quantification of BrdU incorporation. Along pulse-chased replication intermediates from Saccharomyces cerevisiae, we can orient replication tracks and reproduce population-based replication directionality profiles. Additionally, we can map individual initiation and termination events. Thus, FORK-seq reveals the full extent of cell-to-cell heterogeneity in DNA replication.
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FORKseqProtocol_final.pdf (245.71 Ko)
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Figure 1 Forkseq protocol.pdf (899.98 Ko)
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Licence : Domaine public
Origine : Fichiers produits par l'(les) auteur(s)