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 Video

Updated: Nov 26, 2025

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
07:57

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

Published on: May 10, 2022

5.6K

Flash-comet assay.

Erik Bivehed1, Björn Hellman1

  • 1Uppsala University, Department of Pharmaceutical Biosciences, Drug Safety and Toxicology, Uppsala, Sweden.

Methodsx
|December 11, 2020
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

Discovery of Ribosomally Synthesized and Post-translationally Modified Knottins from the Deep-Sea Sponge <i>Stryphnus fortis</i>.

Journal of natural products·2026
Same author

Gel-free capillary zone electrophoresis at acidic pH for micro DNA and RNA analysis.

Talanta·2025
Same author

Visualizing DNA single- and double-strand breaks in the Flash comet assay by DNA polymerase-assisted end-labelling.

Nucleic acids research·2024
Same author

Screening for antibacterial and cytotoxic activities of Sri Lankan marine sponges through microfractionation: Isolation of bromopyrrole alkaloids from Stylissa massa.

PloS one·2024
Same author

DNA integrity under alkaline conditions: An investigation of factors affecting the comet assay.

Mutation research. Genetic toxicology and environmental mutagenesis·2023
Same author

Corrigendum to "Flash-comet: Significantly improved speed and sensitivity of the comet assay through the introduction of lithium-based solutions and a more gentle lysis" [Mutat. Res. - Genet. Toxicol. Environ. Mutagen. 858-860 (2020) 503240].

Mutation research. Genetic toxicology and environmental mutagenesis·2021
Same journal

Facile synthesis of model polystyrene nanoparticles for nanoplastics research.

MethodsX·2026
Same journal

Effectiveness of a posture education program in high school students: A randomized controlled trial protocol.

MethodsX·2026
Same journal

Development and characterization of silicone-based testosterone propionate implants for sustained androgen delivery in juvenile castrated male pigs.

MethodsX·2026
Same journal

Machine learning assisted multi-criteria decision-making approaches for site selection: A systematic review.

MethodsX·2026
Same journal

A systematic analytical framework for multi-source municipal solid waste characterization for energy recovery.

MethodsX·2026
Same journal

Decision tree and reinforcement learning for contextual electricity consumption forecasting in buildings.

MethodsX·2026
See all related articles

The Flash-comet assay significantly speeds up DNA damage detection by using lithium hydroxide (LiOH) for faster unwinding and electrophoresis. This enhanced comet assay method is more sensitive and reduces DNA hydrolysis risk.

Area of Science:

  • Molecular Biology
  • Genotoxicology
  • Biochemistry

Background:

  • The alkaline single-cell gel electrophoresis (SCGE) assay, or comet assay, is a standard method for detecting DNA damage.
  • Conventional protocols involve lengthy unwinding and electrophoresis steps, increasing the risk of artifactual DNA hydrolysis.

Purpose of the Study:

  • To introduce a substantially revised and accelerated alkaline comet assay protocol, termed the Flash-comet assay.
  • To enhance the sensitivity and reduce the time required for DNA strand break and alkali-labile site detection.

Main Methods:

  • The Flash-comet assay utilizes lithium hydroxide (LiOH) instead of sodium hydroxide (NaOH) for alkaline unwinding and electrophoresis.
  • Modified wash and lysis solutions were implemented to prevent DNA alkaline hydrolysis.
Keywords:
DNA-damageGel electrophoresisGenotoxicity testingGentle lysisIn vitro comet assayLow conductivity electrophoresis solutionSingle cell analysis

More Related Videos

Comet Assay to Quantify DNA Damage in FLT3 Mutant-expressing 32D Cells after Exposure to Type I and Type II FLT3 Inhibitors
04:36

Comet Assay to Quantify DNA Damage in FLT3 Mutant-expressing 32D Cells after Exposure to Type I and Type II FLT3 Inhibitors

Published on: October 17, 2025

580
Comet Assay as an Indirect Measure of Systemic Oxidative Stress
08:23

Comet Assay as an Indirect Measure of Systemic Oxidative Stress

Published on: May 22, 2015

29.6K

Related Experiment Videos

Last Updated: Nov 26, 2025

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
07:57

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

Published on: May 10, 2022

5.6K
Comet Assay to Quantify DNA Damage in FLT3 Mutant-expressing 32D Cells after Exposure to Type I and Type II FLT3 Inhibitors
04:36

Comet Assay to Quantify DNA Damage in FLT3 Mutant-expressing 32D Cells after Exposure to Type I and Type II FLT3 Inhibitors

Published on: October 17, 2025

580
Comet Assay as an Indirect Measure of Systemic Oxidative Stress
08:23

Comet Assay as an Indirect Measure of Systemic Oxidative Stress

Published on: May 22, 2015

29.6K
  • Higher electrophoresis voltage (5 V/cm) was applied, drastically reducing unwinding and electrophoresis times.

    • Main Results:

    • The Flash-comet assay achieved significantly shorter unwinding (2.5 min) and electrophoresis (1 min) times compared to conventional protocols.
    • The revised method demonstrated higher sensitivity in detecting DNA strand breaks and alkali-labile sites in cells exposed to H₂O₂ or ionizing radiation.
    • Reduced assay times minimize the risk of laboratory-induced DNA hydrolysis.

    Conclusions:

    • The Flash-comet assay offers a faster and more sensitive alternative for detecting DNA damage.
    • Its reduced run-times and improved sensitivity make it valuable for genotoxicity testing and xenobiotic evaluation.
    • The protocol modifications enhance the reliability of DNA damage assessment by mitigating artifactual hydrolysis.