ROX is a passive reference dye which is not required in machines like Rotor-Gene Q from Qiagen because of the centrifugal rotary design. Each PCR tube aligns with the detection optics during centrifugation in such a manner that the fluorescent signal collected don't have sample to sample variation and also the optical pathway is short between the optics and the sample. This eliminates sample-to-sample variations and edge effects seen in PCR instruments with plate setup like the ABI Real-Time PCR.

For what I understood, ROX is needed to assure that all the wells of a plate emit the same amount of light. ROX is more a check for the photographic apparatus than the chemistry in itself. This because, from what the companies had told me during the years (but probably depends on the machine), a picture is taken of the whole plate, then the individual wells are extracted. Thus, it is important that there are no artifacts in the picture.

Since the rotorgene does not suffer from possible warpings of a plate, there is no need for such a check. Wells are analyzed one by one by the laser beam.

Luigi Marongiu I appreciate your help

ROX is a passive reference dye which is not required in machines like Rotor-Gene Q from Qiagen because of the centrifugal rotary design. Each PCR tube aligns with the detection optics during centrifugation in such a manner that the fluorescent signal collected don't have sample to sample variation and also the optical pathway is short between the optics and the sample. This eliminates sample-to-sample variations and edge effects seen in PCR instruments with plate setup like the ABI Real-Time PCR.

Navonil Gogoi Thank you.