FIGURE 4.
Comparison of RT-PCR assays primed using random (A) or target-specific (B) primers. Both reactions were carried out using the Brilliant 2-step RT-PCR kit. The only difference was that assay A was primed using random nonamers primers (Applied Biosystems), whereas assay B was primed using a target-specific primer (Proligo, Paris, France). In each case, 100 ng of total RNA was subjected to 10-fold (A) or 5-fold (B) serial dilutions and reverse transcribed using standard RT conditions as specified by the manufacturer. One tenth of each cDNA preparation was then included in a PCR assay. dR, baseline-corrected raw fluorescence; dRn, baseline-corrected normalized fluorescence.