Chapter title |
Twelve-Gel Comet Assay Format for Quick Examination of DNA Damage and Repair
|
---|---|
Chapter number | 16 |
Book title |
Fast Detection of DNA Damage
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7187-9_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7185-5, 978-1-4939-7187-9
|
Authors |
Sergey Shaposhnikov, Andrew Collins |
Abstract |
The comet assay (single cell gel electrophoresis) is a sensitive, versatile method for detecting DNA damage in eukaryotic cells. The traditional comet assay format has 1 or 2 gels on a microscope slide, 1 sample per slide, and there is a limit of 40 gels per experiment given the size of a typical electrophoresis tank. To increase throughput, we have designed and tested a system with 12 minigels on one slide, allowing analysis of up to 12 times more samples in one electrophoresis run. The novel comet assay format compares well with the traditional technology. The various steps are suitable for further automation, and the formats can be adapted to fully automated scoring. The new procedures save time at all stages as fewer slides are handled, and the amounts of reagents needed are reduced significantly. This format is particularly useful for testing of numerous genotoxic agents and nanomaterials at different concentrations and on different types of cells; simultaneous analysis of different lesions using a range of enzymes; and analysis of cell extracts for DNA repair activity. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 8 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 2 | 25% |
Student > Postgraduate | 2 | 25% |
Professor > Associate Professor | 1 | 13% |
Student > Doctoral Student | 1 | 13% |
Unknown | 2 | 25% |
Readers by discipline | Count | As % |
---|---|---|
Pharmacology, Toxicology and Pharmaceutical Science | 2 | 25% |
Agricultural and Biological Sciences | 2 | 25% |
Unknown | 4 | 50% |