Hi all,

I have a question regarding calculations using the Pfaffl method.

I have 3 groups per experiment: 1 control group (calibrator) and 2 treatment groups with 6 to 7 biological repeats per group.

Although the averages of the delta Ct values both for the HKG and GOI for the calibrator equal 0 exactly, the averages of the corresponding E^delta Ct s do not equal 1 exactly.

The average of the expression ratios for the calibrator thus ranges from 1 tot 1.3 (not 1 exactly) I did not round off any numbers.

Am I doing the calculations correctly or am I making an error somewhere?

I followed this example:

https://toptipbio.com/pfaffl-method-qpcr/#:~:text=What%20is%20the%20Pfaffl%20method,Nucleic%20Acids%20Research%20in%202001.

Thank you for any suggestions!

Marlies