Does anybody know if I can use the termocycler for real time PCR that doesn't have the adequate instrument to measure the ROX fluorophore with the master mix that have it? I thought that I will have not problems, but when I decided to do my real time PCRs, someone recommended not to use the ROX master mix in this equipment... why!? The most the major problem will be that I will not have the normalization, but will I have any other problems?
The presence of ROX in your mastermix wil not interfere with the MJ Opticon. Unless there are other imcompatibilities you are able to use this mastermix.
The ROX is in the mastermix to normalize when a single CCD camera is used to measure fluoresence. In this case, fluorescent intensity is different when wells are located further from the centre of the plate. The ROX is used to correct for this.
The MJ Opticon has a dedicated detector for each well, so it doesn't need the internal normalization. If you're using standard SYBR green settings, the machine will ignore the ROX signal.
thank you very much for your answers!!!! Knowing what you said before I will work more relax in my following experiments!!!
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