What positive controls are typically included in qPCR and/or qRT-PCR experiments?
It is critical to include appropriate positive controls in a qPCR experiment, in order to determine if false negatives are being detected in the experiment. Positive controls fall into one of two classes. 1. Exogenous positive controls refer to the use of external DNA or RNA carrying a target of interest. If these positive controls are assayed in separate wells/tubes from the experimental sample, they serve as a control for whether or not the reverse transcription and/or PCR reaction conditions are optimal. Additionally, the exogenous DNA or RNA positive controls may be spiked into the experimental sample(s), and assayed in parallel to, or in a multiplex format with, the target of interest. These control reactions assess if the samples contain any components that inhibit reverse transcription and/or PCR. 2. Endogenous positive controls refer to the use of a native target that is present in the experimental sample(s) of interest, but is different from the target under study. These types
Related Questions
- Can the efficiency of the immunoprecipitation (testing the positive and negative controls) be assessed using endpoint PCR instead of qPCR?
- What positive controls are typically included in qPCR and/or qRT-PCR experiments?
- What negative controls are typically included in qPCR and/or qRT-PCR experiments?
