How can I distinguish non-template amplification (NTC) from real products?

The simplest way to tell if non-template amplification is occurring and affecting accurate quantitation is to include control reactions that contain the primers of interest but exclude the target input (with only sample buffer or water added in its place). These no-template control (NTC) reactions may produce amplification products, which will usually be of a different size and/or sequence content than the desired DNA product and will therefore denature with a different melt profile. Comparing melt curves from the different reactions can enable detection of any non-template amplification. Any reactions showing a melt profile similar to the NTC should be removed from the standard curve and quantitation calculations.

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