I am using all the fresh working stocks, water, aerosol tips, changed the working room areas and further more decontaminated the working area with bleach and UV. Still than consistently false positives are shown in no template control. why?
Are you amplifying a human DNA target? If so, it's likely one of your original stocks (not working stocks) of primer, dNTPs, or reaction buffer is contaminated with human DNA as a result of mishandling either by someone in your lab or even by the supplier of the reagent. When we have encountered this problem we started over with fresh, never opened stocks of reaction buffer and dNTPs, and with newly synthesized primers. You can do experiments with other primer pairs to determine which of your reagents is contaminated, but it may not be worth the effort. Buffer stock and primers are cheap. dNTPs are not so cheap, so before tossing them it might be worthwhile to borrow enough from another lab just to see if that solves the problem.
Change stocks and primers.
If you have time make an amplification without DNA and sequencing your product.
You can UV treat water, buffer, and Mg2+. Never do it with dNTPs, primers and Taq DNA pol.
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