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

DNA orientation using specific avidin-ferritin biotin end labelling.

B Theveny, B Revet

    Nucleic Acids Research
    |February 11, 1987
    PubMed
    Summary

    This study presents a new method to determine DNA molecule orientation using biotin labeling and electron microscopy. This technique allows for precise mapping of DNA structures and protein interactions.

    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

    Nucleosome dynamics. Protein and DNA contributions in the chiral transition of the tetrasome, the histone (H3-H4)2 tetramer-DNA particle.

    Journal of molecular biology·1999
    Same author

    Four dimers of lambda repressor bound to two suitably spaced pairs of lambda operators form octamers and DNA loops over large distances.

    Current biology : CB·1999
    Same author

    Isolation and characterization of a protein with high affinity for DNA: the glutamine synthetase of Thermus thermophilus 111.

    Journal of molecular biology·1999
    Same author

    Nucleosome dynamics. II. High flexibility of nucleosome entering and exiting DNAs to positive crossing. An ethidium bromide fluorescence study of mononucleosomes on DNA minicircles.

    Journal of molecular biology·1999
    Same author

    Short unligated sticky ends enable the observation of circularised DNA by atomic force and electron microscopies.

    Nucleic acids research·1998
    Same author

    Interaction of the histone (H3-H4)2 tetramer of the nucleosome with positively supercoiled DNA minicircles: Potential flipping of the protein from a left- to a right-handed superhelical form.

    Proceedings of the National Academy of Sciences of the United States of America·1996

    Area of Science:

    • Molecular Biology
    • Biophysics
    • Biochemistry

    Background:

    • Determining DNA molecule orientation is crucial for understanding DNA structure and function.
    • Existing methods for DNA orientation determination can be complex and time-consuming.

    Purpose of the Study:

    • To develop a rapid and general method for determining DNA molecule orientation.
    • To enable precise mapping and quantification of DNA secondary structures and protein-DNA interactions using electron microscopy.

    Main Methods:

    • End-labeling DNA molecules (phi X174, pBR322, SV40) with a biotin-labeled analog of dTTP (Bio-dUTP).
    • Complexing the Bio-dUTP labeled DNA with Avidin-Ferritin.
    • Imaging the labeled DNA molecules using Electron Microscopy (EM).

    Main Results:

    • Successfully determined the orientation of various DNA molecules.
    • Demonstrated the ability to precisely map DNA secondary structures.
    • Showcased the utility for quantifying protein-DNA interaction sites.

    Conclusions:

    • The developed Bio-dUTP and Avidin-Ferritin labeling method provides an unambiguous way to determine DNA orientation.
    • This technique is rapid, general, and applicable to various DNA molecules for detailed structural and interaction analysis via EM.

    Related Experiment Videos