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What negative controls are typically included in qPCR and/or qRT-PCR experiments?

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What negative controls are typically included in qPCR and/or qRT-PCR experiments?

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The three most common negative controls included in a qPCR and/or qRT-PCR experiment are as follows: 1. No Template Control (NTC) omits any DNA or RNA template from a reaction, and serves as a general control for extraneous nucleic acid contamination. When using SYBR Green chemistry, this also serves as an important control for primer dimer formation. 2. No Reverse Transcriptase Control (NRT) or Minus Reverse Transcriptase Control (MRT) involves carrying out the reverse transcription step of a qRT-PCR experiment in the absence of reverse transcriptase. This control assesses the amount of DNA contamination present in an RNA preparation. 3. No Amplification Control (NAC) omits the DNA polymerase from the PCR reaction. This is a control for background fluorescence that is not a function of the PCR. Such fluorescence is typically attributable to the use of a degraded dual-labeled probe. This control is unnecessary when utilizing SYBR-Green probe chemistries.

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