Duplexing TaqMan Gene Expression Assays

Applied Biosystems TaqMan Gene Expression Assays are easily duplexed, since both predesigned and custom TaqMan Gene Expression Assays are available with either FAM or VIC fluorescent reporter dyes. The most common duplex reaction is combining the target of interest and endogenous control assays in the same well. An endogenous control gene or set of genes is typically used to control for experimental variability due to RNA input and shows expression levels that are relatively constant and moderately abundant across tissues, cell types, and treatment protocols.

Precious samples—when cDNA is limited, duplexing allows more targets to be analyzed since two targets are measured using a single input of cDNA

Cost—run half the number of reactions compared to singleplex qPCR, thereby using half as much master mix and fewer plastic consumables

Precision—data quality is improved because pipetting variability is minimized when the target of interest and endogenous control are measured within the same well

Normalization—use an endogenous control assay in the same well as your target of interest to normalize the expression results and correct for any potential biases caused by the amount of RNA input, reverse transcription efficiency, or sample quality

Learn more about choosing endogenous controls for your experiment ›

Find a predesigned TaqMan Gene Expression Assay ›