Skip to Main content Skip to Navigation

Cartographie pangénomique à haut débit et en molécule unique de la réplication de l'ADN

Abstract : DNA replication is a vital process ensuring accurate conveyance of the genetic information to the daughter cells. In eukaryotic organisms, genome replication is carried out by using multiple start sites, also known as replication origins. In metazoans, the mapping of replication remains challenging. Genome wide mapping of human replication origins performed using sequencing techniques only modestly agree. These existing genome wide approaches use large cell populations that smooth out variability between chromosomal copies that could explain this inconsistency. Thus, to get a better understanding of DNA replication and to uncover the cell-to-cell variability, the development of single molecule techniques is fundamental. DNA combing, a widespread technique used to map DNA replication at a single molecule level, is refractory to automation, forestalling genome-wide analysis. To overcome these impediments, we repurposed an optical DNA mapping device based on microfluidics, the Bionano Genomics Irys system, for High-throughput Optical MApping of Replicating DNA (HOMARD). We typically collect, for a single run, over 34 000 images and more than 63 000 Mbp of DNA. Our new open source tools, that required the adaptation of the provided proprietary software, empower us to simultaneously visualize the intensity profiles of all mapped DNA molecules, check the optical mapping performed and, in particular, see where the replication tracks are located genome-wide at a single molecule level. We demonstrate the robustness of our approach by providing an ultra-high coverage (23,311 x) replication map of bacteriophage DNA in Xenopus egg extracts and the potential of the Irys system for DNA replication and other functional genomic studies apart from its standard use.
Complete list of metadata

Cited literature [240 references]  Display  Hide  Download
Contributor : Abes Star :  Contact
Submitted on : Thursday, October 1, 2020 - 12:01:10 PM
Last modification on : Wednesday, November 17, 2021 - 12:31:57 PM
Long-term archiving on: : Monday, January 4, 2021 - 8:47:15 AM


Version validated by the jury (STAR)


  • HAL Id : tel-02954741, version 1



Nikita Menezes Braganca. Cartographie pangénomique à haut débit et en molécule unique de la réplication de l'ADN. Génomique, Transcriptomique et Protéomique [q-bio.GN]. Université Paris sciences et lettres, 2019. Français. ⟨NNT : 2019PSLEE040⟩. ⟨tel-02954741⟩



Record views


Files downloads