Dear RG-friends,
I would like to check if there are any DNA fragmentations following treatment in primary cells. I know that the comet assay would be best, and since we do not have it at moment, I was looking for an alternative way to check the DNA fragments.
So, can I load cell pellets in a normal agar (0.5-0.7%????) gel? If so, should the pellet be lysed first (for example in SDS+THCl+Glycerol) and then run the electrophoresis? Or, do you have any alternative (simple :-)) way that might suit for my purpose?
Thank you so much in advance for your support. I am looking forward to hearing from you in the near future.
Have a nice day.
Best Reagards,
Mario
Dear Mario, the cell pellet can be used but it needs to be lysed first followed by addition of protease and RNase to avoid any contamination. After that, it can be run on electrophoresis using an appropriate buffer such as TAE or TBE.
Hope it helps.
Good Luck!
Hello
Yes i agree with researchers this is not easy, and use the Triton X-100 better than SDS
Kindly look at in this link may help you
https://www.researchgate.net/post/How_can_I_carry_out_SDS-PAGE_with_whole_cells_Ecoli
Good Luck
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