We performed several PCR assays to detect virulence factor of pathogenic strains, but a faint band amplified on negative control. After this, we have changed all reaction components (one by one) but the band still appearing. Anyone with a similar problem? How did you solved it? Thanks in advance.
Maybe you can try increasing the annealing temperature, that way the primers will have less chances to bind to areas of the genome where the complementarity of the bases is not 100%.
Contamination in PCR can be a frustrating issue, but there are several steps you can take to eliminate it. Here are a few suggestions:
By following these steps, you may be able to identify and eliminate the source of the contamination in your negative control, and obtain reliable results from your PCR assays.
These video playlists might be helpful to you:
https://www.youtube.com/playlist?list=PLzBiAPKi8vONSqjcweFWHMRy13y3484bE
https://www.youtube.com/playlist?list=PLzBiAPKi8vOMheKwb0mu1dcZRaeGUFSlw
https://www.youtube.com/playlist?list=PLzBiAPKi8vOPbE7Db9mKwloe34E8rXXEU
Thanks both for your answers. I´ll follow your kind suggestions.
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