All surfaces in the PCR area should be routinely decontaminated to prevent cross contamination. Use of a DNA decontamination solution, such as DNAzap™, that destroys DNA, is recommended. A “No Template Control” (NTC) should be run to rule out cross contamination of reagents and surfaces. The NTC includes all of the RT-PCR reagents except the RNA template. Typically the RNA is simply substituted with nuclease-free water.
No product should be synthesized in the NTC; if a product is amplified, it indicates that one or more of the RT-PCR reagents is contaminated with the amplicon.