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What negative controls do I need to use for my qPCR experiments?

IDT recommends the following three controls. At the minimum, the no template control is absolutely necessary in all qPCR experiments

  1. No template control – omits the DNA or RNA template from the PCR reaction. This reaction serves as a general control for unwanted nucleic acid contamination or primer dimmer formation that may make the results more difficult to interpret when using SYBR Green chemistry.
  2. No reverse transcriptase control – omits the reverse transcriptase in the reverse transcription step of a qRT-PCR reaction.  The purpose of this control is to assess the amount of DNA contamination present in an RNA preparation.
  3. No amplification control - omits the DNA polymerase from the PCR reaction. This reaction serves as a control for background fluorescence of the PCR which may occur if the dual-labeled probe is degraded.

1 Comment

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