Hi,
I have performed an RT-qPCR for relative quantification. The SOI is frmA and reference gene is topB. I believe there should be no Cq for the control samples labelled as NRT (No-reverse-transcriptase) and NTC (No-template-control). However, there is a Cq for them. Therefore there is genomic contamination.
But my question is this: If there was more double stranded genomic material (either the cDNA or genomic contamination) would there be a lower Cq value for the RT treated? Since there is a high concentration of double stranded material present and the exponential amplification of the amplicon synthesis can be reached before?
-This is comparing the NRT and RT samples for the frmA gene.
I would really appreciate your help if I am not interpreting the data correctly.
Thank you
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