I am performing qPCR for virus absolute quantification with external standards for the virus (say CMV) and also a standard from a seal herpesvirus which I spike in parallel wells containing samples. I quantify the spiked target with a 5-log plasmid curve.
If I want to adjust CMV quantities for inhibitor presence I can use the formula Cq adjusted = Cq sample * ln (1+Espike) / ln(2). Is that correct? What’s the meaning of ln2?
Would it be possible to correct directly linear quantities given by the qPCR platform? The simplest way would be doing
CMV adj = (theoretical number of input copies of spike)/ (measured number of input copies of spike) * CMV measured copy number
How could I adjust further in respect of the efficiencies of both curves? Do you suggest another way to deal with the calculations?
Is there a way to do that with Genex?
Thank you very much for your help!