I am currently doing alkaline comet assays on a HaCaT cell line. I'm fairly new to comet assays. I would like to ask opinion about the result I obtained for my experiment. So I run untreated (picture 1 and 2) and also positive control with H2O2, 200 uM for 30 minutes (picture 3 and 4).
I expect that comet tail should appeared for positive control. But somehow, from the picture, I cannot identify any comet tail.
I would be very thankful if someone can clarify the problem, and whether I’ve done it correctly or vice versa. Thanks!
Have you tried using a lower H2O2 concentration (and shorter incubation time, about 5 minutes)?? I'm positive that's the problem
Jorge Serment
I haven't try that one. I will try it first, hopefully will get a better result.
Thank you for your reply..
Dear s. Zulaikha,
Greetings!!
By looking at the pictures you provided, it seems that most likely your cells are not getting lysed properly. During the lysis step, Please ensure that you keep your sample slides in the lysing solution for a minimum of 2 hours (at 4 degrees Celsius).
Best Wishes,
Samrat
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