Dear Dr Ali, I feel some of the parameters need either review or check such as concentration of agarose, duration of alkaline incubation, time, temp, voltage gradient etc, related to elecrophoresis; one should consider standard reference ie, cells with known amount of specific damage to DNA. Aliquats frozen from single large batch of cells - ether treated and untreated (negative control), treatment with H2O2 / X-ray to induce strand brakes (positive control) for basic assay/photo sensitizers plus light to oxidize guanine positive control.
Consider 'Olive tail moment' and 'tail moment' factors
Dr Y K Lahir
Comet assay is good for detecting overall DNA damage or even oxidative damage. You can also perform methods using anitbodies, such as FACS or ICC/IHC plus fluorescent microscopy. A better way to quantify DNA damage, including identification of its various forms, is via HPLC coupled with tandem MS.
An excellent method and very contrasted and with abundant bibliography in this respect is the study of the 8-hydroxy-deoxyguanosine (8-OH-dG). It is valid for cells, serum and urine. A good choice is, 8-OH-deoxyguanosine concentration (DNA / RNA oxidative damage EIA kit, Cayman Chemical®); 96 plaques wells.
Dear Ahmad Ali,
The SCGE is very good for: 1) By collecting data at the individual cell level allowing for more robust statistical analysis, a large number of cells in the sample (
Please refer the attached file where you can see the applications of Micronucleus Assay, Chromosomal Aberrations and Comet assay.
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