I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big difference between the top and bottom of the gel? The gel was prepared with constant stirring so that EtBr should be distributed evenly.
Hello,
It's just because ethidium bromide is positively charged contrary to DNA. So it migrate toward negative pole in the upper part of the gel.
Hello Berzhan
I agree with Clement. EtBr migrates in the opposite direction to that of DNA because EtBr is positively charged. So you see that the upper half stains more than the lower part of the gel. Uniform staining of the gel can be achieved by staining the gel with EtBr for 10-20 mins after the run is complete and then carry out UV exposure. Also please use fresh TBE buffer for making the gel.
Good Luck.
Berzhan Kurmanov
This is not caused because of EtBr or dye. It is just because of the camera filter in your GelDoc instrument is misplaced and causing the shadow. Try to adjust the focus and this problem will be gone.
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