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Overview
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The mixed isomers of 5(6)-ROX are a fluorescent dye used in RT-PCR methods as an internal reference to determine fluorescence variation that is not associated with the amplification process (plastic of the wells, small differences in concentration or volume, instrument measurements). It produces a constant fluorescence emission signal during the PCR process that is used to normalise the emission produced by the reporter. The fluorescence signal is compatible with most reporters. It is used to label the 5’ end of oligonucleotides as a reporter in the presence of a quencher at the 3’ end (dual labelled probe). During amplification, the dye is cleaved, and the fluorescence increases proportionally with the amount of the specific sequence amplified during the PCR process. The development of the fluorescence signal is therefore specifically related to the amplification of the target sequence.
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