Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

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...
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...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
DNA Replication02:40

DNA Replication

DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication uses a large number of...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Mapping the genetic landscape of the DNA damage response with Cas12a-based combinatorial knockout screens.

bioRxiv : the preprint server for biology·2026
Same author

Replication stress primes a trophectoderm fate in embryonic stem cells.

Cell death discovery·2026
Same author

The DNA helicase HELQ promotes replication fork reversal in coordination with BRCA2- and FANCD2-mediated repair pathways.

Nucleic acids research·2026
Same author

Replicative gaps in DNA damage tolerance, genome instability, and cancer therapy.

Molecular cell·2026
Same author

Targeting the FNIP2-SERCA2b axis improves metabolic and mitochondrial defects in Ataxia Telangiectasia.

Cell death & disease·2026
Same author

West Nile Virus: Epidemiology, Surveillance, and Prophylaxis with a Comparative Insight from Italy and Iran.

Vaccines·2026
Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Jun 1, 2026

Study of the DNA Damage Checkpoint using Xenopus Egg Extracts
10:55

Study of the DNA Damage Checkpoint using Xenopus Egg Extracts

Published on: November 5, 2012

Studying DNA replication fork stability in Xenopus egg extract.

Yoshitami Hashimoto1, Vincenzo Costanzo

  • 1Clare Hall Laboratories, London Research Institute, EN6 3LD, Hertsfordshire, UK. yoshitami.hashimoto@cancer.org.uk

Methods in Molecular Biology (Clifton, N.J.)
|June 11, 2011
PubMed
Summary
This summary is machine-generated.

Replication fork stability is crucial for genome integrity. New Xenopus egg extract assays enable biochemical studies of DNA replication fork stability when encountering DNA damage.

More Related Videos

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
07:18

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis
11:04

A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis

Published on: December 19, 2015

Related Experiment Videos

Last Updated: Jun 1, 2026

Study of the DNA Damage Checkpoint using Xenopus Egg Extracts
10:55

Study of the DNA Damage Checkpoint using Xenopus Egg Extracts

Published on: November 5, 2012

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
07:18

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis
11:04

A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis

Published on: December 19, 2015

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Cell survival and genome stability depend on accurate DNA replication.
  • Understanding DNA replication fork progression is vital, especially when encountering obstacles like DNA damage.
  • Previous research has been limited by the absence of a suitable cell-free system for studying replication in the presence of DNA damage.

Purpose of the Study:

  • To develop and present novel cell-free assays for studying DNA replication fork stability.
  • To investigate the biochemical mechanisms underlying replication fork stability in the presence of DNA damage using a vertebrate system.

Main Methods:

  • Utilizing cell-free extracts from Xenopus laevis eggs.
  • Developing biochemical assays to monitor DNA replication fork progression and stability.
  • Introducing DNA damage or obstacles to mimic in vivo conditions.

Main Results:

  • Successfully established functional cell-free assays for studying DNA replication in Xenopus egg extracts.
  • Demonstrated the utility of these assays in analyzing replication fork stability when encountering various impediments.
  • Provided a platform for detailed biochemical investigation of DNA replication fork dynamics.

Conclusions:

  • The developed Xenopus egg extract system offers a powerful tool for biochemical analysis of DNA replication fork stability.
  • This system facilitates the study of how replication forks respond to DNA damage, advancing our understanding of genome stability maintenance.