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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.

SUBMITTER: Tanner NA

PROVIDER: S-EPMC2651787 | biostudies-literature | 2009 Mar

REPOSITORIES: biostudies-literature

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

Tanner Nathan A NA Loparo Joseph J JJ Hamdan Samir M SM Jergic Slobodan S Dixon Nicholas E NE van Oijen Antoine M AM

Nucleic acids research 20090120 4

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 inhibitor ...[more]

PMID: 19155275

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