Good day, I try to make a pcr nested but I have problems of contamination, I use as a template the negative product of the pcr in the second pcr and gives me positive. I thought that the pollution was in the purification and I eliminated the passage, but still the problem persists. What can I do to avoid contamination of my nested pcr?
Is it possible the primers used in second PCR contaminated?
Have you tried to set up a -ve control for both outer and inner primer pairs?
Apparently it was a mistake of inexperience because after reading I must give a spin to the products before opening them by the aerosols that they can generate and I was not doing. I did it and in the test with only controls the contamination disappeared. Now, I'll try the samples. Thank you
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