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

Quantitative in situ hybridization using initial velocity measurements.

E Cash, M Brahic

    Analytical Biochemistry
    |September 1, 1986
    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

    Circadian disruption and biomarkers of tumor progression in breast cancer patients awaiting surgery.

    Brain, behavior, and immunity·2015
    Same author

    Interleukin 22 is a candidate gene for Tmevp3, a locus controlling Theiler's virus-induced neurological diseases.

    Genetics·2007
    Same author

    Homology between a 173-kb region from mouse chromosome 10, telomeric to the Ifng locus, and human chromosome 12q15.

    Genomics·2001
    Same author

    Viral load increases in SJL/J mice persistently infected by Theiler's virus after inactivation of the beta(2)m gene.

    Journal of virology·2001
    Same author

    Disruption of differentiated functions during viral infection in vivo. V. Mapping of a locus involved in susceptibility of mice to growth hormone deficiency due to persistent lymphocytic choriomeningitis virus infection.

    Virology·2001
    Same author

    Borna disease virus persistent infection activates mitogen-activated protein kinase and blocks neuronal differentiation of PC12 cells.

    The Journal of biological chemistry·2000

    This study introduces a new method for quantifying viral RNA within single cells. Measuring initial hybridization rates offers a more accurate way to count viral genomes compared to saturation methods.

    Area of Science:

    • Molecular Biology
    • Virology
    • Cell Biology

    Background:

    • In situ hybridization is a key technique for quantifying viral RNA at the single-cell level.
    • Existing saturation hybridization methods face challenges in accurately measuring high viral copy numbers.

    Purpose of the Study:

    • To develop an alternative in situ hybridization approach for accurate viral RNA quantification.
    • To establish a method effective in the range of 600 to 60,000 viral genome copies per cell.

    Main Methods:

    • Utilizing initial hybridization rates with low cDNA probe concentration and short incubation times.
    • Measuring autoradiographic grain counts as a proxy for viral genome number.
    • Validating the method for a 7-kb RNA genome.

    Related Experiment Videos

    Main Results:

    • A linear relationship was observed between autoradiographic grain counts and viral genome copy number.
    • The method demonstrated accuracy in the range of 600 to 60,000 copies per cell.
    • This approach overcomes limitations of saturation hybridization at high viral loads.

    Conclusions:

    • The initial hybridization rate method provides a sensitive and accurate alternative for single-cell viral RNA quantification.
    • This technique enhances the ability to study viral dynamics in a wide range of infection levels.