What are the major reasons that results unsuccessful observation of comet tail during comet assay; My sample is fish embryo/ fish larvae. Any body please help me based on the practical experience. Grateful thanks for your kind cooperation.
Is the problem arising in the control group or treated group ? what is your test substance ?
I am doing alkaline assay and problem arising with both treated and control group. My test substances is Japanese medaka (Oryzias latipes) embryos. I am following the methods described in the literature of Dasgupta et al. 2014. Environ Sci Pollut Res. 21:13867–13876. If any one have some explanation regarding the unsuccessful observation of comet tails then please suggest. It will be very helpful for me. Thanks
As already mentioned, the possible causes could be the voltage used or the lysis time. We are used to set the voltage in 1.0V/cm between the electrodes of the electrophoresis vat.
We use red cells of fish blood in our experiments. Comparing with the protocol described by Dasgupta et al. 2014, we use a LMP agarose in lower concentration (0,5%) and we don't add another layer of LMP agarose after removing the first coverslip.
Hope this can help you. Good luck.
Thanks for your kind cooperation @ Prof. Matheus Mantuanelli Robert.
I already found the methods described in the literature of Manzano et al. (2014). I will also try with the methodology described in that paper.
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