RTGreen (the ideal substitute for Sybr Green and EvaGreen) is a very sensitive dye for the detection of double stranded DNA (dsDNA). The dye is a green fluorescent nucleic acid dye with features that make it useful for non-specific detection of amplification in realtime qPCR experiments. Compared with the widely used SYBR Green I, RTGreen dye is generally less inhibitory toward PCR and less likely to cause nonspecific amplification , RTGreen dye can be used at a much higher dye concentration than SYBR Green I, resulting in more robust PCR signal. The PCR reaction can be monitored using our existing optical setting for SYBR Green I or FAM on any commercial real-time PCR cycler.