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

Progress towards single-molecule DNA sequencing: a one color demonstration.

James H Werner1, Hong Cai, James H Jett

  • 1Bioscience Division, Los Alamos National Laboratory, Mail Stop J586, Los Alamos, NM 87545, USA. jwerner@lanl.gov

Journal of Biotechnology
|April 2, 2003
PubMed
Summary

This study demonstrates single-molecule DNA sequencing feasibility by detecting fluorescently labeled nucleotides released during DNA digestion. This method allows for individual reading of DNA sequences on multiple fragments for consensus building.

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

Symmetry-Driven Assembly of a Penta-Shell Keplerate Cuprofullerene for Metallofullerene Frameworks.

Angewandte Chemie (International ed. in English)·2023
Same author

Hikikomori: A perspective from bibliometric analysis.

Psychiatry and clinical neurosciences·2023
Same author

Fabrication of a composite 3D-printed titanium alloy combined with controlled in situ drug release to prevent osteosarcoma recurrence.

Materials today. Bio·2023
Same author

Human alveolar macrophage response to Mycobacterium tuberculosis: immune characteristics underlying large inter-individual variability.

Research square·2023
Same author

Extensive Neumann type of pemphigus vegetans.

Indian journal of dermatology, venereology and leprology·2023
Same author

A new tractable method for generating human alveolar macrophage-like cells <i>in vitro</i> to study lung inflammatory processes and diseases.

mBio·2023

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Biophysics

Background:

  • DNA sequencing is crucial for genetic research and diagnostics.
  • Current sequencing methods have limitations in resolution and throughput.
  • Single-molecule detection offers potential for higher accuracy and novel approaches.

Purpose of the Study:

  • To demonstrate the feasibility of single-molecule DNA sequencing.
  • To develop a method for detecting individual nucleotides during DNA enzymatic cleavage.
  • To explore a new approach for generating consensus DNA sequences.

Main Methods:

  • Utilizing ultra-sensitive flow cytometry for single-molecule detection.
  • Employing fluorescently labeled nucleotides (tetramethyl rhodamine labeled dUMP) within DNA fragments.

Related Experiment Videos

  • Observing the processive digestion of DNA strands by Exonuclease I.

    • Main Results:

    • Successfully detected single molecules of fluorescently labeled nucleotides released during DNA digestion.
    • Demonstrated that the digestion of a single DNA strand by a single exonuclease is discernible.
    • Achieved high efficiency in single-molecule detection via laser-induced fluorescence.

    Conclusions:

    • The study validates the feasibility of single-molecule DNA sequencing.
    • This technique provides a practical method for obtaining consensus sequences from multiple DNA fragments.
    • Highlights a novel approach for high-resolution DNA analysis.