I would like to perform the micronucleus and comet assays in our cells after exposure to the mixtures we are investigating. In literature there a lot of protocols with big differences in reagent concentrations and types, lysis conditions, exposure time, and so on.
Someone can recommend me some validated protocols with suggestion to perform these assays?
Kind regards.
Hi Diego,
I do not know anything about micronuclei assay but run couple of experiments with comet assay. I used kit from Trevigen, specifically alkaline version. In my hands their protocol did not ask for a lot of optimization. Here is the manual. Good luck.
I also have protocol for comet assay and while trying to perform it in my sample I had a numerous difficulties. Here is official protocol made by a group of researchers and we flowed it in details. One major problem was lysis and we had quite big problems, the time for this step is suggested to be from 1 day - few weeks. After a vast number of experiments we have came to 24h to be suitable, but again you need to try it on your own.
Good luck.
Please consult publications by Professor Tolga Çavaş (Cavas in Latin alphabet) of Biology Department Uludag University, Bursa Turkey:
http://biyoloji.uludag.edu.tr/eng/ctolga.html
and Professor Goran Klobucar of Croatia:
https://www.pmf.unizg.hr/biol/en/goran.klobucar