What is needed for performing delta delta Ct relative quantitation on the Rotor-Gene Q?

In delta delta Ct analysis, each sample input amount is normalized by comparing with a normalizing gene. These values are then normalized relative to a calibrator. A calibrator can e.g. be a wild-type control, untreated control or a time-zero sample.

A standard curve for each run is not necessary once the assay has been validated.

The amplification efficiency should be identical for both the gene of interest and normalizing gene.

It is important to clearly define samples in the “edit samples’ window. Sample names should be identical when defining pages where the gene of interest and normalizing gene are analyzed.

Refer to section 7-6-4 of the Rotor-Gene Q User Manual for instruction or contact QIAGEN Technical Service for further information.