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

A bead-based method for multiplexed identification and quantitation of DNA sequences using flow cytometry.

A Spiro1, M Lowe, D Brown

  • 1Physics Department, Loyola College in Maryland, Baltimore, Maryland 21210, USA.

Applied and Environmental Microbiology
|September 30, 2000
PubMed
Summary

A novel bead-based assay using flow cytometry offers superior DNA sequence discrimination and accurate quantification for environmental samples. This method enhances the analysis of microbial communities in complex mixtures.

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

Every Person Counts in a Fair Transition to Net Zero: A UK Food Lens Towards Safeguarding Against Nutritional Vulnerability.

Nutrition bulletin·2025
Same author

Meat and the future of sustainable diets-Challenges and opportunities.

Nutrition bulletin·2024
Same author

Temporary screw epiphyseodesis of the first metatarsal for correction of juvenile hallux valgus.

Journal of children's orthopaedics·2018
Same author

Vitamin D: An overview of vitamin D status and intake in Europe.

Nutrition bulletin·2015
Same author

Circulating irisin levels and coronary heart disease: association with future acute coronary syndrome and major adverse cardiovascular events.

International journal of obesity (2005)·2014
Same author

Calcium homeostasis influences radiological fracture healing in postmenopausal women.

Archives of orthopaedic and trauma surgery·2012

Area of Science:

  • Molecular Biology
  • Environmental Science
  • Analytical Chemistry

Background:

  • Identifying specific DNA sequences in complex environmental samples is crucial for microbial ecology and environmental monitoring.
  • Existing methods like DNA microarrays can face limitations in sequence discrimination and accurate quantification.

Purpose of the Study:

  • To develop and evaluate a novel multiplexed, bead-based nucleic acid hybridization method for identifying and quantifying DNA sequences.
  • To assess the performance of this bead assay in terms of sequence discrimination, sensitivity, and detection limits.

Main Methods:

  • Utilized 5.6-microm polystyrene beads with attached oligomer capture probes and embedded fluorophores for multiplexed detection.
  • Employed flow cytometry to analyze fluorescently labeled cDNA sequences captured from environmental samples.

Related Experiment Videos

  • Developed a fluorescence standard for accurate quantification and compared results with DNA microarray analysis.

    • Main Results:

    • The bead-based method demonstrated significantly superior sequence discrimination compared to DNA microarrays.
    • Accurate quantification of target DNA sequences, obtained from the 16S/23S intergenic spacer region of groundwater microorganisms, was achieved.
    • The assay's resolution, sensitivity, and lower detection limit for measuring DNA abundances were determined.

    Conclusions:

    • The developed multiplexed bead assay provides a powerful tool for analyzing heterogeneous DNA mixtures, particularly in environmental microbiology.
    • This method offers enhanced sequence discrimination and facilitates precise quantification, outperforming traditional DNA microarray techniques for specific applications.