Real-time single-molecule observation of rolling-circle DNA replication

We present a simple technique for visualizing replication of individual DNA molecules in real time. By attaching a rolling-circle substrate to a TIRF microscope-mounted flow chamber, we are able to monitor the progression of single-DNA synthesis events and accurately measure rates and processivities...

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Detalles Bibliográficos
Autores principales: Tanner, Nathan A., Loparo, Joseph J., Hamdan, Samir M., Jergic, Slobodan, Dixon, Nicholas E., van Oijen, Antoine M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2009
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2651787/
https://www.ncbi.nlm.nih.gov/pubmed/19155275
http://dx.doi.org/10.1093/nar/gkp006
Descripción
Sumario:We present a simple technique for visualizing replication of individual DNA molecules in real time. By attaching a rolling-circle substrate to a TIRF microscope-mounted flow chamber, we are able to monitor the progression of single-DNA synthesis events and accurately measure rates and processivities of single T7 and Escherichia coli replisomes as they replicate DNA. This method allows for rapid and precise characterization of the kinetics of DNA synthesis and the effects of replication inhibitors.

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