Attached are three photos of example results I see after I run the comet assay on plant leaf tissues. I have been following Gichner's comet assay protocol for plant tissue involving slicing plants in buffer so the nuclei fall out. These examples are after a 15 min. unwinding in NaOH buffer followed by 30 min. of electrophoresis at 26V, and a 5 min ethidium bromide stain. Each time I run the comet assay, I get many odd shapes of many different sizes, but they never appear as the nice, orderly circles that are published in the literature. I'm not convinced that the items in my photos are actually comets. Can anyone suggest what I'm doing wrong?
These images are very different from the general cell comets. I think, you should standardize the whole protocol in your system of work. Again, making single cell suspension is also a vital task which should be done carefully.
As Plant cells look different from that of animal cells, therefore the appearance of cell comets may be different from that of animal cells.
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