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

Nathan A. Tanner; Joseph J. Loparo; Samir M. Hamdan; Slobodan Jergic; Nicholas E. Dixon; Antoine M. van Oijen

doi:10.1093/nar/gkp006

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

Nathan A. Tanner, Joseph J. Loparo, Samir M. Hamdan, Slobodan Jergic, Nicholas E. Dixon, Antoine M. van Oijen^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

68 Scopus citations

Abstract

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.

Original language	English (US)
Article number	e27
Journal	NUCLEIC ACIDS RESEARCH
Volume	37
Issue number	4
DOIs	https://doi.org/10.1093/nar/gkp006
State	Published - 2009

Bibliographical note

Funding Information:
This work was supported by the National Institutes of Health (GM077248 to A.v.O.); Australian Research Council (to N.E.D.); and Jane Coffin Childs Foundation (to J.J.L.). Funding for open access charge: National Institutes of Health (GM077248).

ASJC Scopus subject areas

Genetics

Access to Document

10.1093/nar/gkp006

Cite this

@article{f99c8d41a2d448948d3b93dca7cccf53,

title = "Real-time single-molecule observation of rolling-circle DNA replication",

abstract = "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.",

author = "Tanner, {Nathan A.} and Loparo, {Joseph J.} and Hamdan, {Samir M.} and Slobodan Jergic and Dixon, {Nicholas E.} and {van Oijen}, {Antoine M.}",

note = "Funding Information: This work was supported by the National Institutes of Health (GM077248 to A.v.O.); Australian Research Council (to N.E.D.); and Jane Coffin Childs Foundation (to J.J.L.). Funding for open access charge: National Institutes of Health (GM077248).",

year = "2009",

doi = "10.1093/nar/gkp006",

language = "English (US)",

volume = "37",

journal = "NUCLEIC ACIDS RESEARCH",

issn = "0305-1048",

publisher = "OXFORD UNIV PRESS",

number = "4",

}

TY - JOUR

T1 - Real-time single-molecule observation of rolling-circle DNA replication

AU - Tanner, Nathan A.

AU - Loparo, Joseph J.

AU - Hamdan, Samir M.

AU - Jergic, Slobodan

AU - Dixon, Nicholas E.

AU - van Oijen, Antoine M.

N1 - Funding Information: This work was supported by the National Institutes of Health (GM077248 to A.v.O.); Australian Research Council (to N.E.D.); and Jane Coffin Childs Foundation (to J.J.L.). Funding for open access charge: National Institutes of Health (GM077248).

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=62549122138&partnerID=8YFLogxK

U2 - 10.1093/nar/gkp006

DO - 10.1093/nar/gkp006

M3 - Article

C2 - 19155275

AN - SCOPUS:62549122138

SN - 0305-1048

VL - 37

JO - NUCLEIC ACIDS RESEARCH

JF - NUCLEIC ACIDS RESEARCH

IS - 4

M1 - e27

ER -

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

Abstract

Bibliographical note

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this