EvaGreen® dye is a green fluorescent nucleic acid dye with features that make the dye useful for several applications including qPCR and DNA melt curve analysis, real-time monitoring of thermophilic helicase-dependent amplification (tHDA), and capillary gel electrophoresis. The DNA-bound dye has excitation and emission spectra very close to those of fluorescein (FAM) or SYBR® Green I, making the dye readily compatible with instruments equipped with the 488 nm argon laser or any visible light excitation with wavelength in the region. EvaGreen® dye is extremely stable both thermally and hydrolytically, providing convenience during routine handling. The dye is essentially nonfluorescent by itself, but becomes highly fluorescent upon binding to dsDNA. EvaGreen® dye is nonmutagenic and noncytotoxic by being completely impermeable to cell membranes, making it a great alternative to SYBR® Green I, which enters cell rapidly and is known to be a powerful mutation-enhancer (Ohta, et al. (2001). Mutation Research, 492(1), 91-97).

The unique properties of EvaGreen® dye have made it particularly useful in quantitative real-time PCR (qPCR) applications. Compared with the widely used SYBR® Green I, EvaGreen® dye is generally less inhibitory toward PCR and less likely to cause nonspecific amplification. As a result, EvaGreen® dye can be used at a much higher dye concentration than SYBR® Green I, resulting in a high resolution signal for droplet digital PCR, real-time PCR and melt curve analysis.