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
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author Tanner, Nathan A.
Loparo, Joseph J.
Hamdan, Samir M.
Jergic, Slobodan
Dixon, Nicholas E.
van Oijen, Antoine M.
author_facet Tanner, Nathan A.
Loparo, Joseph J.
Hamdan, Samir M.
Jergic, Slobodan
Dixon, Nicholas E.
van Oijen, Antoine M.
author_sort Tanner, Nathan A.
collection PubMed
description 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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spelling pubmed-26517872009-03-13 Real-time single-molecule observation of rolling-circle DNA replication Tanner, Nathan A. Loparo, Joseph J. Hamdan, Samir M. Jergic, Slobodan Dixon, Nicholas E. van Oijen, Antoine M. Nucleic Acids Res Methods Online 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. Oxford University Press 2009-03 2009-01-20 /pmc/articles/PMC2651787/ /pubmed/19155275 http://dx.doi.org/10.1093/nar/gkp006 Text en © 2009 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Tanner, Nathan A.
Loparo, Joseph J.
Hamdan, Samir M.
Jergic, Slobodan
Dixon, Nicholas E.
van Oijen, Antoine M.
Real-time single-molecule observation of rolling-circle DNA replication
title Real-time single-molecule observation of rolling-circle DNA replication
title_full Real-time single-molecule observation of rolling-circle DNA replication
title_fullStr Real-time single-molecule observation of rolling-circle DNA replication
title_full_unstemmed Real-time single-molecule observation of rolling-circle DNA replication
title_short Real-time single-molecule observation of rolling-circle DNA replication
title_sort real-time single-molecule observation of rolling-circle dna replication
topic Methods Online
url 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
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