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

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

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

Author(s) -

Nathan A. Tanner,

Joseph J. Loparo,

Samir M. Hamdan,

Slobodan Jergic,

Nicholas E. Dixon,

Antoine M. van Oijen

Publication year - 2009

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkp006

Subject(s) - replisome , biology , rolling circle replication , dna replication , dna , replication (statistics) , total internal reflection fluorescence microscope , biophysics , eukaryotic dna replication , biological system , genetics , virology , membrane

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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