The Qubit 2.0 Fluorometer High-Sensitivity kit for DNA quantification relies on a 2-point standard curve to generate your data.
The appendix indicates the Qubit uses a "modified Hill Plot" using your 2-point standard curve to generate the regression equation to backfit your sample data.
I found a paper on making Hill Plots but it's way too math-heavy for me to follow.
I've used the kit components to make a standard curve with various points between the 2 already provided with the kit and have observed a non-linear relationship between the DNA input and the Green RFU values. Obviously, a simple linear regression is not appropriate for backfitting the data, and yet I don't understand how the fluorometer can be doing anything else since it only uses a 2-point curve.
My question, how can you generate a regression equation that is nonlinear using only 2 points on a standard curve?