I want to perform a comet assay on mouse neuron cells. Because it is a lot easier for work and for experiment if I can freeze mouse brain and do the comet assay later, I am curious if the cells from tissue that has been frozen can be used in comet test. Or it is obligatory to use always live cells?
The cells in this assay are eventually lysed before electrophoresis so it seems logical that I can use frozen samples.
as u r going to study DNA fragmentation, there is no problem using frozen tissue. DNA is very stable and does not alter much.
you can do comet assay for frozen cells, there are published reports. my suggestion is after electrophoresis acquire image as early as possible to get good result.
If I can use frozen samples then why in some experiments they determine cell viability? And only perform comet assay on viable cells. Because if you freeze and then unfreeze cells they will be dead for sure.
please go through the paper,
http://www.sciencedirect.com/science/article/pii/S1383571803003255#
Best conditions for Comet assay, using fresh samples. However, if you do not test in time, you can store samples at -80 ° C freezer. The samples should not be kept for more than two or three months in the freezer.
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