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EvaGreen® Dye, 20X in Water

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.

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Product 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 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 (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) 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.

See our ready-to-use Forget-Me-Not™ EvaGreen® qPCR master mix as well as our master mix with 2-color tracking for sensitive, competitively-priced qPCR reagents. We also offer Thiazole Green, 1000X in DMSO, which is identical to SYBR® Green I.

EvaGreen dye and applications are covered by granted US and international patents.


Low PCR inhibition

Exhibits 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 qPCR and melt curve analysis signals

Nonmutagenic and noncytotoxic

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

Compatible with fast PCR protocols

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, chemical 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; withstands repeated freeze-thaw cycles

Spectrally similar to SYBR® Green I

Compatible with all major brand qPCR instruments and enzyme systems


Mao, et al. Characterization of EvaGreen Dye and the implication of its physicochemical properties for qPCR applications. BMC Biotechnology 7, 76 (2007).

EvaGreen in droplet digital PCR

1. Lab Chip (2010), doi: 10.1039/c004521g

2. The Royal Society of Chemistry (2014), doi: 10.1039/c3an02334f

3. Analytical Chemistry (2014), doi: 10.1021/ac403843j

4. Analytical Chemistry (2013),

5. The Journal of American Society (2011),

6. Analytical Chemistry (2010), doi: 10.1021/ac902510u

7. Analytical Chemistry (2013), doi: 10.1021/ac403061n

8. Nucleic Acids Research (2013), doi:10.1093/nar/gkt684

Download a list of Selected EvaGreen References.


You may also like…

    Thiazole Green, 10,000X in DMSO

    Thiazole Green, which is structurally identical to SYBR® Green I Nucleic Acid Gel Stain, is one of the most sensitive stains available for detecting double-stranded DNA (dsDNA) in agarose, polyacrylamide gels and qPCR.

    Forget-Me-Not™ EvaGreen® qPCR Master Mix (2-Color Tracking)

    Forget-Me-Not™ EvaGreen® is a high performance qPCR master mix with 2-color reaction component tracking system available for a fraction of the cost of similar products.

    GelRed® Nucleic Acid Gel Stain

    GelRed® is an ultra sensitive, extremely stable and environmentally safe fluorescent nucleic acid dye designed to replace the highly toxic ethidium bromide (EB) for staining dsDNA, ssDNA or RNA in agarose gels or polyacrylamide gels. GelRed® is far more sensitive than EB without requiring a destaining step.

    GelGreen® Nucleic Acid Gel Stain

    GelGreen® is a sensitive, stable and environmentally safe green fluorescent nucleic acid dye designed to stain either dsDNA, ssDNA or RNA in agarose gels. GelGreen® is far more sensitive than SYBR Safe.

    Ready-to-Use 100 bp DNA Ladder

    100 bp DNA Ladder is suitable for sizing linear double-stranded DNA fragments from 100 bp to 1500 bp. The 500 bp and 1,500 bp bands contain more DNA to provide internal orientation.

    EvaEZ™ Fluorometric Polymerase Activity Assay Kit

    EvaEZ™ Fluorometric Polymerase Activity Assay Kit provides an easy and accurate way to determine the activity of a nucleic acid polymerase without using radioisotopes. It contains EvaGreen® dye together with a primed template, dNTPs, and MgCl2 in a Tris buffer system. Available without magnesium by special order.

Support & Faq

EvaGreen® dye and Master Mixes

EvaGreen® dye is spectrally similar to FAM or SYBR® Green I, which means no change in optical settings is required for using an EvaGreen®-based master mix. However, there are several differences between the dyes.

Lower background fluorescence: EvaGreen® dye has less background than SYBR® Green I due to its novel “release-on-demand” DNA-binding mechanism.

Brighter and non-inhibitory:  EvaGreen® is less inhibitory in the PCR reaction than SYBR® Green, permitting the use of saturating dye concentration for maximal signal and better high-resolution DNA melt analysis.

Safer and more environmentally friendly: EvaGreen® dye is the first and only PCR dye to date designed to be environmentally safe.

Dye stability: EvaGreen® dye is very stable both during storage and under PCR conditions. SYBR® Green I, on the other hand, is known to degrade following multiple freeze-thaw cycles and under PCR conditions. Moreover, decomposed SYBR® Green I is reported to be even more inhibitory to PCR.

Digital PCR: EvaGreen® dye is currently the only qPCR dye to be used in droplet digital PCR (ddPCR).

Category: EvaGreen® dye and Master Mixes

← FAQs

Biotium offers three different EvaGreen® dye-based qPCR master mixes:

  • Forget-Me-Not™ qPCR Master Mix
  • Fast EvaGreen® qPCR Master Mix
  • Fast Plus EvaGreen® qPCR Master Mix

All three master mixes contain our high-quality EvaGreen® DNA-binding dye and our fast-activating chemically-modified hotstart enzyme, Cheetah™ Taq.

Forget-Me-Not™ qPCR Master Mix is our newest master mix formulation. For most purposes this is the master mix that we would recommend, as it has many benefits: best sensitivity and signal, blue tracking dyes in the master mix and template to reduce pipetting errors, and low cost. Fast EvaGreen® qPCR Master Mix is a sensitive, general use master mix that does not contain blue tracking dyes. Fast Plus EvaGreen® qPCR Master Mix has a slightly different formulation than the regular Fast EvaGreen® qPCR Master Mix, and was designed for more challenging samples.

Category: EvaGreen® dye and Master Mixes

← FAQs

Both of these qPCR master mixes contain Cheetah™ Taq (our chemically modified hotstart DNA polymerase), buffer and dNTPs.

The Fast EvaGreen® Master Mixes also contain the DNA-binding dye EvaGreen®, and are used for fluorescent dye-based qPCR. The only thing the user needs to supply are primers and template.

The Fast Probe Master Mix  is formulated for conducting real-time PCR using an oligo-based probe, such as a TaqMan™ probe, an MGB probe or an AllGlo™ probe. The user needs to supply primers, probe and template.

Category: EvaGreen® dye and Master Mixes

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