Ensuring that qPCR is quantitative, accurate, and reproducible requires thoughtful assay design, setup, and optimization. During design, each of the MMolecular RT-qPCR assays follows stringent criteria to ensure optimal performance.
Each MMolecular assay meets the following criteria:
-PCR efficiency 2.0 +/- 0.2 (equals 100 +/- 10 %).
-Cq of highest cDNA concentration <= 34.
-Linear dynamic range of at least 3 logs.
-High amplification specificity, with no side products in gel analysis.
-Sigmoidal amplification curve.
-Fluorescence intensity of amplification curves between 5 and 50 fluorescence units.