Description
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.
Features
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Exhibits much less PCR inhibition than SYBR® Green I via a smart “release-on-demand” DNA-binding technology
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Low PCR inhibition of the dye permits a higher dye concentration to be used for much greater qPCR and melt curve analysis signals
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- Nonmutagenic and noncytotoxic
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Nonmutagenic and noncytotoxic by standard Ames test; completely impermeable to cell membranes
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- Compatible with fast PCR protocols
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Minimal interference to PCR makes it possible to significantly shorten the chain extension time
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- Compatible with multiplex PCR
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No dye migration from amplicon to amplicon when used at the recommended concentration
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- Unsurpassed thermal stability, chemical stability and photostability
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No detectable dye decomposition in PCR buffer at 95-100°C for 48 hours; highly stable under either alkaline or acidic condition; withstands repeated freeze-thaw cycles
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- Spectrally similar to SYBR® Green I
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Compatible with all major brand qPCR instruments and enzyme systems |
Storage/Handling
Store at -20°C. Protect from light.
Product is stable for at least 12 months when stored correctly.
Product may be shipped at ambient temperature without reducing performance. Please place at the recommended storage conditions upon receipt of the product.
EvaGreen® dye and applications are covered under US patent nos. 7,803,943 and 7,776,567 and pending international patents.
