What is the difference between a relative quantification assay and an absolute quantification assay?
A relative quantification (also known as comparative quantification) assay quantifies changes in gene expression relative to a reference gene and reference sample. An absolute quantification assay uses a standard curve of known quantities to determine the quantity of unknown samples. Relative quantification results report fold changes in expression relative to the reference gene and reference sample. Absolute quantification results are reported as an absolute quantity (copies, ng, pg, etc.) extrapolated from the standard curve.
A relative quantification (also known as comparative quantification) assay quantifies changes in gene expression relative to a reference gene and reference sample. An absolute quantification assay uses a standard curve of known quantities to determine the quantity of unknown samples. Relative quantification results report fold changes in expression relative to the reference gene and reference sample. Absolute quantification results are reported as an absolute quantity (copies, ng, pg, etc.) extrapolated from the standard curve. • What is the difference between one-step and two-step real time RT-qPCR The difference between one-step and two-step real time RT-qPCR lies mainly in the reverse transcription step. In one-step RT-qPCR, a short reverse transcription (5–30 minute) reaction is followed by a PCR reaction in a single tube. In two-step RT-qPCR the reverse transcription reaction takes place in a separate tube. Each method has advantages and disadvantages, depending on the application
