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How do I evaluate the performance of a real-time PCR reaction?
Posted November 25, 2020

Answer

The performance of real-time PCR can be evaluated on the following criteria:

  • Dynamic Range: this determines the efficiency of a PCR reaction and can be accomplished by performing a 5-log dilution series. A slope of -3.3 ± 10% is ideal as this indicates a PCR efficiency of 100%.
  • Precision: defines the closeness of agreement between measurements performed under the same conditions. This is determined by calculating the standard deviation, with a standard deviation <0.167 being ideal.
  • Sensitivity: this describes the lowest amount of starting material that can be detected or measured.