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

Mutational spectrometry without phenotypic selection: human mitochondrial DNA

K Khrapko1, H Coller, P André

  • 1Division of Toxicology, Center for Environmental Health Sciences, MIT, Cambridge, MA 02139, USA.

Nucleic Acids Research
|February 15, 1997
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

Percutaneous cholecystostomy in critically ill patients with acute cholecystitis: complications and late outcome.

Clinical radiology·2014
Same author

The evaluation of a novel technique to treat saphenous vein incompetence: preclinical animal study to examine safety and efficacy of a new vein occlusion device.

Phlebology·2012
Same author

Influence of column configuration on performance in high efficiency liquid chromatography.

Analytical chemistry·2012
Same author

Conjugation reactions in the preparations of quantum dot-based immunoluminescent probes for analysis of proteins by capillary electrophoresis.

Analytical and bioanalytical chemistry·2011
Same author

Age-related gene-specific changes of A-to-I mRNA editing in the human brain.

Mechanisms of ageing and development·2010
Same author

Single molecule PCR in mtDNA mutational analysis: Genuine mutations vs. damage bypass-derived artifacts.

Methods (San Diego, Calif.)·2008

Researchers developed a new method to detect rare mitochondrial DNA point mutations. This technique allows for the observation of mutations at very low frequencies in biological samples.

Area of Science:

  • Molecular Biology
  • Genetics

Background:

  • Point mutations in mitochondrial DNA (mtDNA) are implicated in various diseases.
  • Detecting rare mutations at low frequencies is crucial for understanding disease mechanisms.

Purpose of the Study:

  • To develop a sensitive method for observing point mutations in human mtDNA.
  • To identify mutation hotspots and their origins in cellular and tissue samples.

Main Methods:

  • Separation of mutant and non-mutant DNA sequences based on melting temperatures.
  • High-fidelity DNA amplification to increase the number of mutant DNA copies.
  • Analysis of point mutations in specific regions of the mitochondrial genome.

Main Results:

  • A method capable of detecting point mutations at fractions of 10^-6 or higher was established.

Related Experiment Videos

  • Hotspot point mutations within a 100-base pair region of the mtDNA were observed in cultured cells and human tissues.
  • Nineteen mutants were isolated, with fractions ranging from 4x10^-4 to the limit of detection.
  • Distinction between true mutants and potential artifacts (contamination, PCR errors, DNA adducts) was performed.

    • Conclusions:

    • The developed technology is effective for observing the spectrum of point mutations in human mtDNA.
    • The method serves as a tool for investigating the primary causes of mtDNA mutations.
    • Some observed mutations may originate from mismatch intermediates or endogenous DNA adducts, in addition to double-stranded point mutations.