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The present QC-PCR assay differs from others described previously by enabling accurate analysis of multiple samples with a single set of standards over a wide range instead of requiring multiple assays for the titration of each sample [1, 2].
In the present QC-PCR, 1000 copies of the competitor DNA, the value at the middle of the logarithmic dynamic range, was added to a dilution series of target DNA for establishing a calibration curve.
In developing the present QC-PCR assay, we constructed the competitor DNA by means of a splice overlap extension PCR method.
A limitation of this QC-PCR is that the RT efficiency of the sample RNA was not controlled in the presence of an internal RNA standard.