Thermo Scientific ROX dye is added to the qPCR reactions to serve as an internal reference for normalization of the fluorescent signal when using real-time instruments that can detect ROX. ROX allows for the correction of well-to-well variation appearing due to pipetting inaccuracies and fluorescence fluctuations. It can be added to an entire 2X master mix tube or to an individual reaction mixture (for recommended amounts of ROX Solution for a specific real-time instrument refer to the qPCR manual).
ROX does not participate in PCR and does not interfere with the specificity or sensitivity of qPCR assays.
ROX Solution contains ROX dye as a 50 µM aqueous solution at pH 7.6. The excitation maximum and emission maximum of ROX are at 580 nm and 621 nm, respectively.