Absolute quantification and relative quantification are two ways that are used to calculate the results of a qPCR experiment. There are a few significant differences between the two.
Basis of differentiation | Absolute Quantification | Relative Quantification |
What it determines | Expression levels in absolute numbers of copies | Fold changes in expression between two samples |
Known/Unknown template amounts | The precise amount of the message or template used for the curve is known. The absolute quantification standard curve provides the final answer. | Although the template is known to contain the message of interest in high abundance, its absolute amount may or may not be known. |
What the process involves | Unknowns are quantified based on a known quantity. A standard curve is first created. Then the unknowns are compared to the standard curve and a value is extrapolated. | Changes in gene expression in a given sample are analyzed relative to another reference sample such as an untreated control sample. |
How the answer is determined | The standard curve provides the final answer. | The calibration curve result for the target gene is normalized to that of a housekeeping gene in the same sample. The normalized numbers are then compared between samples to obtain a fold change. |