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ROX is used as passive reference dye to compensate for non-PCR related well to well variations in the fluorescence. Variations in fluorescence can occur e.g. due to well to well variation in light intensity, which depends on the optical construction of the PCR machine or due to pipetting variations. The ROX fluorescence does not change during the course of the PCR reaction but provide a stable baseline to which samples are normalized. The excitation and emission of the reference dye are 584nm and 612nm, respectively.
The ROX concentration to be used depends on the Real Time PCR instrument or, more precisely, on the filter available. On earlier Real Time PCR machines there was no filter matching precisely to the ROX fluorescence. Therefore, high ROX concentrations are needed with these machines. Some newer machines do not need any internal standard to correct for light intensity. Nevertheless, an internal standard might still be useful to correct for pipetting variations.
This dye has been qualified for use on instruments like the ABI PRISM® 7700, One-Step System, RotorGene and many others.
Please consult the user manual of the real time PCR instrument with regard to appropriate levels of ROX. Ready to use qPCR Master Mixes with High ROX (500nM) >, Low ROX (50nM) > or without ROX > are also available from Genaxxon.
Concentration: 200µM ROX dissolved in 20mM Tris/HCl, pH8.5, 0.1mM EDTA, 0.01% Tween® 20
Delivered as 3 x 200µL
application:For realtime PCR instruments that need a ROX concentration of 500nM (High ROX) the dilution factor is 1:400 (0.12µL per 50µL) PCR reaction (3µL per 1.25mL 2-time Mastermix).For realtime PCR instruments that need a ROX concentration of 50nM (Low ROX) the dilution factor is 1:4000 (0.012µL per 50µL) PCR reaction (0.3µL per 1.25mL 2-time Mastermix).
Sicherheits Hinweise / Safety
Klassifizierungen / Classificationeclass-Nr: 32-16-04-03
Dokumente - Protokolle - Downloads
Here you will find information and further literature on ROX Passive Reference Dye - 600 µL. For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: email@example.com or phone: +49 731 3608 123.