When I performed Dot blot assay I always see a "ring signal" in the sonicated DNA sample (positive ctrl).
What do you think, how can I avoid this phenomenon?
Hello Agnes,
this issue was broadly discussed previously on research gate.
Hope this link help:
https://www.researchgate.net/post/Ring_pattern_observed_in_dot_blot
I will try to the spotting step using lower volume and in our previous experiment we applied ctrl samples where the buffers contained different salt (MgCl2, NaCl). In these cases ring pattern was not detected. Salt could help..
50 mM NaCl was added to the sample and ring pattern dissappered. Volume of sample spotting onto the membrane was 1,5 microliter. The fluorescent detection isn't perfect, probably lack of UV fixation step. I repeat the experimnet.
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