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Related Concept Videos

The DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...

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Updated: Jun 19, 2026

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy
15:57

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

Published on: October 9, 2009

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy.

Nathan A Tanner1, Joseph J Loparo, Antoine M van Oijen

  • 1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School.

Journal of Visualized Experiments : Jove
|October 13, 2009
PubMed
Summary
This summary is machine-generated.

We developed a simple fluorescence microscopy method to observe DNA replication at the single-molecule level in real-time. This technique precisely measures replication rates and processivity using a forked DNA template and laminar flow.

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Last Updated: Jun 19, 2026

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy
15:57

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Published on: October 9, 2009

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Published on: October 27, 2011

Area of Science:

  • Molecular Biology
  • Biophysics
  • Microscopy

Background:

  • Understanding DNA replication is crucial for cell division and genetic stability.
  • Observing DNA replication at the single-molecule level provides detailed insights into the process.
  • Existing methods may lack real-time capabilities or require complex setups.

Purpose of the Study:

  • To develop a simple, real-time fluorescence microscopy method for observing DNA replication.
  • To enable precise measurement of DNA replication rate and processivity at the single-molecule level.

Main Methods:

  • A forked DNA template was immobilized on a functionalized glass coverslip.
  • Replication proteins and nucleotides were introduced to initiate DNA synthesis.
  • The growing double-strand DNA (dsDNA) product was visualized using an intercalating dye under laminar flow.
  • The position of the growing DNA end was tracked in real-time using fluorescence microscopy.

Main Results:

  • The method allows real-time observation of DNA replication at the single-molecule level.
  • Precise determination of DNA replication rates was achieved by measuring the growing DNA end position.
  • The length of completed DNA products provided information on replication processivity.

Conclusions:

  • This fluorescence microscopy-based method offers a simple and rapid approach to study DNA replication.
  • The technique facilitates real-time analysis of replication dynamics, including rate and processivity.
  • The described method requires only a standard fluorescence microscope and a sensitive camera.