How do I determine the amplification efficiency of my qPCR assay?

To determine the amplification efficiency of your qPCR assay, you must first make five serial dilutions of your target molecule in 2-fold, 5-fold, and 10-fold serial dilutions.

Use each serial dilution in separate real-time reactions to determine their threshold cycle values (Ct values).

Plot the Ct values obtained on a logarithmic scale along with corresponding concentrations.

Generate a linear regression curve through the data points and calculate the slope of the trend line.

Amplification efficiency of your qPCR assay can be calculated using the equation E = -1+10^(-1/slope).