We recommend the following 3 negative controls:

-The “no template control” (NTC) is an absolute requirement for all qPCR experiments. An NTC omits DNA from the PCR. This reaction serves as a general control for unwanted nucleic acid contamination.

-A no reverse transcriptase control (–RT) omits the reverse transcriptase in the reverse transcription step of a RT-qPCR. The purpose of this control is to assess the amount of genomic DNA contamination present in an RNA preparation. Perform this control for any assay that might amplify
genomic DNA.

-A no amplification control omits the DNA polymerase from the PCR. This reaction serves as a control for background fluorescence of the PCR assay and probe stability.