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

39.2K
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...
39.2K
The Replisome03:01

The Replisome

37.3K
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
37.3K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

6.1K
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,...
6.1K
Homologous Recombination02:31

Homologous Recombination

60.0K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
60.0K
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

5.2K
The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of...
5.2K
DNA Replication02:40

DNA Replication

56.3K
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...
56.3K

You might also read

Related Articles

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

Sort by
Same author

Proteolytic cleavage activates the mitochondrial isoform of TOP3A.

Nucleic acids research·2025
Same author

Author Correction: Ribonucleotide incorporation into mitochondrial DNA drives inflammation.

Nature·2025
Same author

Ribonucleotide incorporation into mitochondrial DNA drives inflammation.

Nature·2025
Same author

Small molecules restore mutant mitochondrial DNA polymerase activity.

Nature·2025
Same author

Mechanistic basis of atypical TERT promoter mutations.

Nature communications·2024
Same author

The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics.

Nucleic acids research·2024

Related Experiment Video

Updated: Nov 29, 2025

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

40.4K

In Vitro Analysis of mtDNA Replication.

Jay P Uhler1, Maria Falkenberg2

  • 1Department of Medical Biochemistry and Cell Biology, University of Gothenburg, Gothenburg, Sweden. jennifer.uhler@medkem.gu.se.

Methods in Molecular Biology (Clifton, N.J.)
|November 24, 2020
PubMed
Summary

Researchers detail how to reconstitute human mitochondrial DNA replication in vitro. This study focuses on differentially replicating the leading heavy-strand and lagging light-strand for cellular energy production.

Keywords:
DNA polymeraseIn vitroMitochondriaReplicationmtDNA

More Related Videos

Author Spotlight: High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution
10:47

Author Spotlight: High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution

Published on: May 5, 2023

4.1K
Visualizing Single-molecule DNA Replication with Fluorescence Microscopy
15:57

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

Published on: October 9, 2009

22.9K

Related Experiment Videos

Last Updated: Nov 29, 2025

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

40.4K
Author Spotlight: High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution
10:47

Author Spotlight: High-Throughput Image-Based Quantification of Mitochondrial DNA Synthesis and Distribution

Published on: May 5, 2023

4.1K
Visualizing Single-molecule DNA Replication with Fluorescence Microscopy
15:57

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

Published on: October 9, 2009

22.9K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Human mitochondrial DNA (mtDNA) is crucial for cellular energy production.
  • Replication of the mtDNA genome is performed by a specialized protein machinery, including DNA polymerase γ.
  • The current model suggests asynchronous replication of mtDNA strands: leading heavy-strand first, then lagging light-strand.

Purpose of the Study:

  • To provide detailed methods for reconstituting mitochondrial DNA replication in vitro.
  • To describe the differential reconstitution of leading- and lagging-strand replication.

Main Methods:

  • Utilizing purified recombinant replication proteins.
  • Employing synthetic DNA templates for in vitro studies.
  • Developing protocols for differential strand replication.

Main Results:

  • Successful reconstitution of in vitro mitochondrial DNA replication.
  • Demonstration of differential replication capabilities for leading and lagging strands.
  • Detailed protocols for researchers to follow.

Conclusions:

  • The study provides a reproducible in vitro system for studying mitochondrial DNA replication.
  • This methodology allows for the independent investigation of leading- and lagging-strand synthesis.
  • Advances understanding of the mechanisms governing mitochondrial genome replication and cellular energy production.