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Article Dans Une Revue Methods Année : 2012

Genome-scale identification of active DNA replication origins

C Cayrou
Philippe Coulombe
Etienne Danis
  • Fonction : Auteur
Marcel Méchali
  • Fonction : Auteur
  • PersonId : 912051

Résumé

Understanding the nature of DNA replication origins in metazoan is quite challenging. In the absence of a genetic assay like in yeast, methods were devised based on the DNA structure, the visualization or quantification of the first nascent strands that are synthesized at origins, or on the localization of origin binding proteins. The purification and quantification of RNA-primed nascent DNA at origins during initiation of DNA synthesis is the most exhaustive and precise method to map active replication origins at present. We have upgraded this method to the level of reproducibility and enrichment required for genome-wide analyses by microarrays or deep sequencing. We detail here the protocol and the controls required at the different steps.

Domaines

Génétique

Dates et versions

hal-00733286 , version 1 (18-09-2012)

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Citer

C Cayrou, Damien Gregoire, Philippe Coulombe, Etienne Danis, Marcel Méchali. Genome-scale identification of active DNA replication origins. Methods, 2012, 57 (2), pp.158-164. ⟨10.1016/j.ymeth.2012.06.015⟩. ⟨hal-00733286⟩
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