What is the difference between Absolute Quantification and Relative Quantification in qPCR, using the standard curve approach?
Absolute Quantification determines expression levels in absolute numbers of copies. Relative Quantification determines fold changes in expression between two samples. In absolute quantification, the precise amount of the message or template used for the curve is known. In relative quantification, the template is simply known to contain the message of interest in high abundance, but its absolute amount is not necessarily known. Unknowns are compared to either standard curve and a value is extrapolated. The absolute quantification standard curve provides the final answer. The relative quantification calibration curve result for the gene of interest is normalized to that of a housekeeping gene in the same sample, and then the normalized numbers are compared between samples to obtain a fold change.
Related Questions
- Can you provide the general basis of the assay kits and how to calculate concentrations from the standard curve and sample data?
- What is the difference between Absolute Quantification and Relative Quantification in qPCR, using the standard curve approach?
- How do I run a melting curve or dissociation stage at the end of a Relative Quantification (RQ) run?
