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

Mitochondrial DNA depletion analysis by pseudogene ratioing.

Russell H Swerdlow1, Gerard T Redpath, Daniel R Binder

  • 1Department of Neurology, University of Virginia Health System, Box 394, Charlottesville, VA 22908, USA. rhs7e@virginia.edu

Journal of Neuroscience Methods
|August 25, 2005
PubMed
Summary

This study introduces a novel mtDNA pseudogene ratioing technique to confirm mitochondrial genome depletion in rho(0) cell lines. This method offers a reliable alternative to traditional PCR or hybridization for assessing mitochondrial DNA copy number.

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

Patient cerebral organoids capture Alzheimer's disease proteomic biomarkers and drug targets.

bioRxiv : the preprint server for biology·2026
Same author

Evaluating the causal effect of mitochondrial dysfunction on Alzheimer's disease and Parkinson's disease using polygenic risk scores and Mendelian randomization.

Alzheimer's & dementia : the journal of the Alzheimer's Association·2026
Same author

An aging hallmark, Alzheimer's disease, and <i>APOE</i> nexus.

Journal of Alzheimer's disease : JAD·2026
Same author

Limbic-predominant neuroimaging correlates of plasma p-Tau217 in preclinical and clinical Alzheimer's disease.

Frontiers in aging neuroscience·2026
Same author

Downward bias in the association between APOE and Alzheimer's disease using prevalent and by-proxy disease sampling in the All of Us research program.

BMC medical genomics·2026
Same author

Structural signature of plasma proteins classifies the status of Alzheimer's disease.

Nature aging·2026

Area of Science:

  • Cell Biology
  • Genetics

Background:

  • Mitochondrial DNA (mtDNA) depletion is crucial for creating rho(0) cell lines.
  • Current methods like hybridization and PCR for assessing mtDNA depletion rely on negative results, which can be ambiguous.

Purpose of the Study:

  • To introduce and validate a novel mtDNA pseudogene ratioing technique for confirming rho(0) cell line status.
  • To provide a more definitive method for assessing mitochondrial DNA depletion and copy number.

Main Methods:

  • Utilized ethidium bromide to induce mtDNA depletion in U251 human glioma cells.
  • Employed primers targeting both mtDNA and a nuclear DNA-embedded mtDNA pseudogene (mtDNApsi) for PCR amplification.
  • Generated and genotyped plasmid clones from PCR products to quantify mtDNApsi relative to mtDNA.

Related Experiment Videos

Main Results:

  • All 62 genotyped plasmid clones originated from the mtDNApsi template, indicating successful mtDNA depletion.
  • Established with 95% confidence that the cell line contains less than one copy of mtDNA per 10 cells.
  • Demonstrated that the mtDNApsi ratioing technique does not depend on the interpretation of negative results.

Conclusions:

  • The mtDNA pseudogene ratioing technique provides a robust and quantitative method for confirming rho(0) status.
  • This adjunct method enhances the reliability of assessing mitochondrial DNA depletion and copy number compared to traditional approaches.