Methods and related aspects for quantitative polymerase chain reaction to determine fractional abundance

A method of processing a grain sample to detect the fractional abundance of a transgenic trait, comprising:

• Grinding and mixing the grain sample,
• Extracting and purifying DNA from the grain sample,
• Running said purified DNA sample in a qPCR reaction, said qPCR reaction comprising at least two different fluorescent probes that competitively bind to the same DNA locus, wherein said DNA locus is indicative of the presence or absence of a transgenic trait,
• Generating results of the qPCR reaction that characterize the competitive binding levels of each of the at least two different fluorescent probes during each cycle of the qPCR reaction, and
• Interpreting said results with one or more of a machine learning model, ratiometric regression algorithm, or best fit model to determine the fractional abundance of the transgenic trait.