What is the current premier assay for monitoring DNA damage, break formation and/or repair in cell culture? Is it immunofluorescence based (e.g. fish, antibody). Or are there commercial assays that exist. Any insight would be greatly appreciated.
Single cell gel electrophoresis (SCGE; comet assay) is a widely-used method due to its technical simplicity and sensitivity.
Please see the attached file:
https://www.nature.com/articles/s41598-018-20995-w
Hello Dane
Besides comet assay, and immunofluorescence in which phosphorylated H2AX is detected at sites of DNA damage by using specific antibody, there are other methods of monitoring DNA damage. Please refer to the research article below.
Oncol Lett. 2017 Jun, 13(6): 3982-3988.
doi: 10.3892/o1.2017.6002
Best Wishes.
Hello, Dane
Please You look at the following guidlines:
Guidelines for DNA recombination and repair studies: Cellular assays of DNA repair pathways
http://microbialcell.com/researcharticles/2019a-klein-microbial-cell/
http://microbialcell.com/wordpress/wp-content/uploads/2019/01/2019A-Klein-Microbial-Cell.pdf
MOLECULAR BIOLOGY ASSAYS TO DETECT DNA BREAKS AND REPAIR
We have used the TUNEL assay many times to quantify DNA damage, read the description this could be useful for you.
https://www.promega.de/en/products/cell-health-assays/apoptosis-assays/deadend-fluorometric-tunel-system/?catNum=G3250
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