ZellX^® One-Step Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR) Kit grants efficient cDNA synthesis and quantitative/real-time PCR in a single tube. The PCR Master Mix provided in the kit contains a green fluorescent reporter dye plus all the reagents (except primers and RNA template) needed for running real-time PCR reactions. In addition, a separate Reverse Transcriptase mix that comprises a balanced mixture of both Reverse Transcriptase and RNase Inhibitor is provided. As an internal reference, the kit contains high concentrations of carboxy-X-rhodamine (ROX^TM). Being independent from the amount of DNA template, the fluorescence signal of ROX^TM is not influenced by the PCR reactions, and therefore can assist in the normalization of the reporter-dye signal during data analysis. The appropriate level of ROX^TM depends on the real-time PCR instrument (Contact your instrument manufacturer for details). For low levels of ROX^TM use our Dye-based one-step RT-qPCR kit with low carboxy-X-rhodamine (Cat NO. ZX-22109-100/200/400).

RT-PCR is used to amplify double-stranded DNA from single-stranded RNA templates. In the RT step the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA) complementary to the RNA template. In the first cycle of the PCR, Taq DNA polymerase synthesizes DNA molecules complementary to the cDNA, thus generating a double-stranded DNA template. During subsequent cycles, the DNA polymerase exponentially amplifies the double-stranded DNA template.