Dear friends,
I could set up micronucleus assay on cell line. but I have a big problem in positive control. I tried vast ranges of H2O2 doses for possitive control(even I used the standard data obtained from MTT assay) but in some dose I couldn't find micronuclei and in other doses the H2o2 killed the cell...I am really confused now..may I ask you give me a guide or advice to solve my problem?
Dear Helena,
Thanks for your nice reply..I will try your advice and I will share my result with you..
Best Regards.
Dear Zahra,
I used doxorubicin for positive control in micronucleous assay in human bladder cancer cells (http://www.ncbi.nlm.nih.gov/pubmed/24625788).
Good luck in the experiments
André
Hi Zahra,
I have not performed micronucleus assay but when I used H2O2 as a positive control in another genetic toxicology assay (in vitro comet) the same sort of thing happened. It is suspected that hydrogen peroxide causes oxidative damage that can sometimes be repaired and therefore does not give a positive genotoxic result!
If you just google H2O2 DNA repair you should find more information on this. So yes I agree that you should try a different positive control... Possibly MMS or MMC as per the OECD guidelines just like Helena suggested.
Good Luck!
Erin
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