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 dye is essentially nonfluorescent by itself, but becomes highly fluorescent upon binding to dsDNA. The DNA-bound dye has excitation and emission spectra very close to those of fluorescein (FAM) or SYBR®, Green I, making it 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. EvaGreen®, dye is nonmutagenic and noncytotoxic because it is impermeable to cell membranes, unlike SYBR®, Green I, which enters cell rapidly and is known to be a powerful mutation-enhancer. The unique properties of EvaGreen®, dye have made it particularly useful in quantitative real-time PCR (qPCR) applications.