EvaGreen^®dye is a green fluorescent nucleic acid dye with features that make the dye useful for several applications including qPCR, high-resolution DNA melt curve analysis (HRM)¹, real-time monitoring of thermophilic helicase-dependent amplification (tHDA), routine solution DNA quantification and capillary gel electrophoresis. The DNA-bound dye has excitation and emission spectra very close to those of fluorescein (FAM) or SYBR^®dye 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, unlike SYBR Green I, which enters cell rapidly and is known to be a powerful mutation-enhancer (Ohta, et el. Mutat. Res. 492, 91(2001)).

The unique properties of EvaGreen dye have made it particularly useful in quantitative real-time PCR (qPCR) application. 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 more robust PCR signal.

We offer four EvaGreen products: 1) Fast EvaGreen Master Mix for qPCR and HRM (cat#31003), 2) EvaGreen dye, 20X in water (cat#31000), 3) EvaGreen, qPCR 2X Basic Mix (cat#31001), and 4) EvaGreen, qPCR 2X Basic Mix HS (for hotstart Taq) (cat#31004). The Fast Master Mix is suitable for both qPCR and high resolution melt curve (HRM) analysis.

Features:

• Very Little PCR inhibition

Exhibit much less PCR inhibition than SYBR Green I via a smart "release-on-demand" DNA-binding technology

• Highly Sensitive

Low PCR inhibition of the dye permits a higher dye concentration to be used for much greater fluorescent signal and high-resolution melt curve analysis (HRM)

• Nonmutagenic and noncytotoxic

Nonmutagenic and noncytotoxic by standard Ames test; completely impermeable to cell membranes

• Compatible with Fast PCR protocol

Minimal interference to PCR makes it possible to significantly shorten the chain extension time

• Compatible with multiplex PCR

No dye migration from amplicon to amplicon when used at the recommended concentration

• Unsurpassed Thermal Stability, Hydrolytical Stability and Photostability

No detectable dye decomposition in PCR buffer at 95-100°C for 48 hours; highly stable under either alkaline or acidic condition; withstand repeated freeze-thaw cycles

• Spectrally similar to SYBR Green I

Compatible with all major brand qPCR instruments and enzyme systems