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 Experiment Videos

Mapping replication fork direction by leading strand analysis

M I Aladjem1, G M Wahl

  • 1Gene Expression Laboratory, Salk Institute, San Diego, California 92037, USA.

Methods (San Diego, Calif.)
|January 27, 1998
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Patents and literature.

Applied biochemistry and biotechnology·2013
Same author

Egg dispersal in the acanthocephalan Acanthocephalus dirus: field data.

The Journal of parasitology·2012
Same author

Functional analysis and consequences of Mdm2 E3 ligase inhibition in human tumor cells.

Oncogene·2012
Same author

How does leflunomide modulate the immune response in rheumatoid arthritis?

BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy·2007
Same author

c-Myc, genome instability, and tumorigenesis: the devil is in the details.

Current topics in microbiology and immunology·2006
Same author

Mouse bites dogma: how mouse models are changing our views of how P53 is regulated in vivo.

Cell death and differentiation·2006
Same journal

An accessible, absorbance-based plate reader assay to assess cumulative exposure of blood plasma & serum to thawed conditions.

Methods (San Diego, Calif.)·2026
Same journal

EC-isHCR: A rapid method for in situ hybridization chain reaction in diverse animal samples.

Methods (San Diego, Calif.)·2026
Same journal

Single-Molecule methods to investigate mechanisms of transcription by RNA polymerase of Mycobacterium tuberculosis.

Methods (San Diego, Calif.)·2026
Same journal

Detection and sequencing of Usutu virus during mosquito surveillance: Use of multiple assays and techniques for identification at low levels.

Methods (San Diego, Calif.)·2026
Same journal

Experimental validation of an AI-driven digital healthcare platform for oral health behavior and plaque assessment among vietnamese children.

Methods (San Diego, Calif.)·2026
Same journal

Zeta potential: An efficient and cost-effective alternative for investigating cell-surface interactions.

Methods (San Diego, Calif.)·2026
See all related articles

This study details a method to determine DNA replication direction using leading strand isolation. This technique helps map DNA replication origins and fork patterns, especially in complex genomic regions.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA replication is a fundamental biological process.
  • Understanding replication fork direction is crucial for genome stability and regulation.
  • Existing methods for mapping replication origins can be limited.

Purpose of the Study:

  • To describe and validate a method for determining replication fork polarity.
  • To provide a protocol for mapping DNA replication initiation sites.
  • To offer guidance on interpreting results from leading strand analysis.

Main Methods:

  • Isolation of newly replicated DNA strands in the presence of emetine to inhibit Okazaki fragment synthesis.
  • Detection of leading strands using single-stranded probes hybridized to cloned genomic templates.

Related Experiment Videos

  • Quantitative hybridization to determine the direction of DNA synthesis.

    • Main Results:

    • The leading strand method allows for the determination of replication fork movement direction.
    • This technique is effective for mapping initiation sites in various cell lines and chromosomal loci.
    • The study reviews experimental details and provides controls to mitigate potential pitfalls.

    Conclusions:

    • The leading strand method is a valuable tool for mapping DNA replication initiation and fork patterns.
    • Careful interpretation is necessary due to reliance on quantitative hybridization and potential variations in efficiency.
    • The protocol facilitates accurate analysis of replication dynamics.