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Real-Time Single-Molecule Observation of Rolling-Circle DNA Replication

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2009

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Oxford University Press
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Tanner, Nathan A., Joseph J. Loparo, Samir M. Hamdan, Slobodan Jergic, Nicholas E. Dixon, and Antoine M. van Oijen. 2009. Real-time single-molecule observation of rolling-circle DNA replication. Nucleic Acids Research 37(4): e27.

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.

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