How to check the primer efficiency for real time PCR ?

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Jochen Wilhelm

The documentation from Applied Biosystems is ok.

Katie A Burnette

You set up a "standard curve" of at least 5 concentrations of your template. You use the slope of the graph of ct value vs. log of copy number to calculate the efficiency. Check out Thermo Fisher's website for more information.

https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-learning-center/real-time-pcr-basics/efficiency-real-time-pcr-qpcr.html#curve

Amir Hossein Mohseni

I suguest use Primer Blast online software of NCBI website along gene runner offine software.

Hamza Yaiche

You can use Primer Blast online software of NCBI website.

Mohammed Omar Baba sheikh

I suggest use use positive sample or positive control for that primer set

Md. Mostafijur Rahman

I agree with @Katie A Burnette. You can also use the melt curve data for different concentrations of template to check whether the primer binding is affected in different concentrations or not.

Md. Zafaryab

Thank u all. I have check suggestion given by Dr. Brunette.