adjusting data with spike quantitation

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adjusting data with spike quantitation

Postby rociohache » Tue Oct 30, 2012 12:30 pm

Hello!
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!

Rocio
rociohache
 
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