Scenario
A qPCR target amplifies in both the reverse-transcribed sample and the no-RT control.
The example focuses on deciding whether the signal can still be interpreted or whether sample preparation must be reviewed first.
Inputs to confirm
Ct values for RT-positive and no-RT reactions.
Primer design context, especially whether primers span exon junctions when applicable.
Whether DNase treatment or genomic DNA removal was performed.
How a researcher would use the result
Compare no-RT signal to the true sample signal before calculating fold change.
If the no-RT control is close to the sample Ct, interpret expression cautiously and review RNA cleanup or primer design.
Common failure points
Ignoring no-RT amplification because the sample Ct looks strong.
Using a reference gene without checking genomic DNA sensitivity.
Treating DNase treatment as guaranteed rather than verified.