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 dual-fluorescence reporter system for high-throughput clone characterization and selection by cell sorting.

Juno Choe1, Haiwei H Guo, Ger van den Engh

  • 1Institute for Systems Biology 1441 North 34th Street, Seattle, WA 98103-8904, USA.

Nucleic Acids Research
|March 16, 2005
PubMed
Summary

This study introduces a novel dual-fluorescence reporter system for efficient DNA sequence isolation. It uses green fluorescent protein (GFP) and Discosoma red fluorescent protein (DsRed) to identify and sort cells containing desired DNA inserts using high-speed flow cytometry.

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

Viral tag and grow: a scalable approach to capture and characterize infectious virus-host pairs.

ISME communications·2023
Same author

Species-targeted sorting and cultivation of commensal bacteria from the gut microbiome using flow cytometry under anaerobic conditions.

Microbiome·2022
Same author

Left Atrial Volume as a Biomarker of Atrial Fibrillation at Routine Chest CT: Deep Learning Approach.

Radiology. Cardiothoracic imaging·2021
Same author

Rituximab Versus Mycophenolate in the Treatment of Recalcitrant Connective Tissue Disease-Associated Interstitial Lung Disease.

ACR open rheumatology·2020
Same author

Loss of Filamentous Multicellularity in <i>Cyanobacteria</i>: the Extremophile <i>Gloeocapsopsis</i> sp. Strain UTEX B3054 Retained Multicellular Features at the Genomic and Behavioral Levels.

Journal of bacteriology·2020
Same author

Thoracic Radiologists' Versus Computer Scientists' Perspectives on the Future of Artificial Intelligence in Radiology.

Journal of thoracic imaging·2019

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Molecular biology relies on isolating specific DNA sequences.
  • Flow cytometry offers high-speed cell analysis and sorting.
  • Single fluorescent reporters have limitations due to expression variability.

Purpose of the Study:

  • To develop a novel dual-fluorescence reporter system to overcome single reporter limitations.
  • To improve the efficiency of clone characterization and DNA sequence isolation.
  • To validate a new reporter system for molecular biology applications.

Main Methods:

  • Developed a dual-reporter system using green fluorescent protein (GFP) and Discosoma red fluorescent protein (DsRed).
  • Utilized high-speed cell sorting based on altered green to red fluorescence ratios.

Related Experiment Videos

  • Validated the system through shotgun library construction, PCR amplification, and DNA sequencing.

    • Main Results:

    • The dual-reporter system successfully identified cells with foreign DNA inserts by altered fluorescence.
    • High-speed cell sorting effectively separated cells containing desired inserts.
    • Validated cloned sequences from sorted bacterial cells.

    Conclusions:

    • Dual-fluorescence reporting coupled with high-speed cell sorting is a more efficient alternative to traditional clone isolation methods.
    • The system is robust, adaptable for biosensor assays, and amenable to miniaturization.
    • This method enhances DNA sequence isolation in molecular biology.