Blank qPCR Master Mix (2x) is a universal solution for quantitative real-time PCR and two-step real- time RT-PCR and can be used on most real-time PCR cyclers available. The master mix does not contain any dye and allows the user to choose a dye and optimize the dye concentration. The master mix contains Perpetual Taq DNA Polymerase, optimized reaction buffer and dNTPs (dTTP is partially replaced with dUTP).
The use of ROX passive reference dye is necessary for all real-time PCR cyclers from Applied Biosystems and optional for cyclers from Stratagene. ROX compensates for variations of fluorescent signal between wells due to slight differences in reaction volume and fluorescence fluctuations. ROX is not involved in PCR reaction, has a different emission spectrum than SYBR Green I and does not interfere with real-time PCR on any instrument. Refer to the table below to determine the recommended amount of ROX required for a specific PCR cycler.