NZYSupreme qPCR Green Master Mix (2x), ROX is an optimized and highly efficient reaction mixture developed for real-time PCR. This master mix was engineered with a hot-start enzyme control mechanism to provide the highest detection sensitivity. In addition, the latest developments in PCR enhancers have been incorporated in the NZYSupreme qPCR Green Master Mix, ROX, including buffer chemistry and incorporation of highly robust engineered enzymes. These combinations guarantee that NZYSupreme qPCR Green Master Mix (2x), ROX delivers ultrasensitivity coupled with highly reproducible and fast real-time PCR protocols. It was designed to amplify targets for accurate gene expression analysis. The master mix is provided as a 2x reaction mixture that contains all components necessary for real-time PCR, including a green intercalating dye, dNTPs, stabilizers and enhancers. The presence of ROX reference dye in the master mix enables to increase confidence in data analysis, since it allows to normalize non-PCR-related fluctuations in fluorescence. Despite most real-time PCR instruments that can read ROX dye allow users to run experiments and analyse data without ROX, the inclusion of this internal passive reference dye prevents data misinterpretation and allows to detect and diagnose errors. NZYSupreme qPCR Green Master Mix (2x), ROX is ready-to-use and only requires primers and DNA template addition. It is optimized for intercalating green dye detection on different instruments.