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 Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Tuberculosis research in South Africa over the past 30 years: From bench to bedside.

South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde·2020
Same author

Generation and characterization of thiol-deficient Mycobacterium tuberculosis mutants.

Scientific data·2018
Same author

Proteomic analysis reveals that sulfamethoxazole induces oxidative stress in M. tuberculosis.

Tuberculosis (Edinburgh, Scotland)·2018
Same author

Antigen-specific interferon-gamma release is decreased following the single intradermal comparative cervical skin test in African buffaloes (Syncerus caffer).

Veterinary immunology and immunopathology·2018
Same author

Detection of tuberculosis by automatic cough sound analysis.

Physiological measurement·2018
Same author

Seroprevalence of Mycobacterium bovis infection in warthogs (Phacochoerus africanus) in bovine tuberculosis-endemic regions of South Africa.

Transboundary and emerging diseases·2018

Related Experiment Video

Updated: Jun 3, 2026

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
12:20

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses

Published on: December 29, 2015

Plasmid preparation on sephacryl s1000.

P D Van Helden1, E G Hoal

  • 1MRC Unit for Molecular and Cellular Cardiology, Department of Medical Physiology and Biochemistry, University of Stellenbosch Medical School, Tygerberg, South Africa.

Methods in Molecular Biology (Clifton, N.J.)
|March 23, 2011
PubMed
Summary

Producing pure plasmid DNA for recombinant DNA research is essential. Current methods involve costly and time-consuming ultracentrifugation, potentially yielding unusable DNA.

Area of Science:

  • Molecular Biology
  • Biotechnology

Background:

  • Pure plasmid DNA is critical for recombinant DNA research.
  • Existing plasmid purification methods involve bacterial growth, lysis, and centrifugation.
  • Ultracentrifugation with cesium chloride and ethidium bromide is a common but expensive step.

Purpose of the Study:

  • To highlight the limitations of current plasmid purification techniques.
  • To emphasize the cost and time associated with ultracentrifugation.
  • To address the issue of enzyme inhibition due to incomplete reagent extraction.

Main Methods:

  • Bacterial growth and lysis.
  • Low-speed centrifugation to remove cellular debris.
  • Ultracentrifugation in cesium chloride and ethidium bromide for DNA separation.

More Related Videos

Production and Visualization of Bacterial Spheroplasts and Protoplasts to Characterize Antimicrobial Peptide Localization
10:13

Production and Visualization of Bacterial Spheroplasts and Protoplasts to Characterize Antimicrobial Peptide Localization

Published on: August 11, 2018

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures
06:53

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures

Published on: January 5, 2011

Related Experiment Videos

Last Updated: Jun 3, 2026

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
12:20

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses

Published on: December 29, 2015

Production and Visualization of Bacterial Spheroplasts and Protoplasts to Characterize Antimicrobial Peptide Localization
10:13

Production and Visualization of Bacterial Spheroplasts and Protoplasts to Characterize Antimicrobial Peptide Localization

Published on: August 11, 2018

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures
06:53

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures

Published on: January 5, 2011

  • Plasmid DNA recovery and ethidium bromide/cesium chloride extraction.

    • Main Results:

    • Ultracentrifugation and subsequent extraction are expensive and time-consuming.
    • Specialized equipment like ultracentrifuges and rotors are required.
    • Incomplete ethidium bromide removal can inhibit downstream enzymatic manipulation of plasmid DNA.

    Conclusions:

    • Current plasmid purification protocols present significant economic and practical challenges.
    • The ultracentrifugation step is a major bottleneck in terms of cost, time, and equipment.
    • Ensuring complete removal of purification reagents is vital for the functional integrity of purified plasmid DNA.