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Bright, sensitive and environmentally safe; ideal for qPCR and many other applications
2-color or 1-color tracking allows you to spot pipetting errors at a glance
Suitable for all fluorescent probe-based PCR methods including TaqMan® or Molecular Beacons
High performance master mix for chip-based digital PCR (cdPCR) platforms.
Chemnically modified hot-start taq; no activity at RT and activates in only 2 minutes
Easy, accurate, and non-radioactive nucleic acid polymerase activity assay
dNTPs, ROX reference dye, hot-start/warm-start enzyme modification kits, and more


EvaGreen® dyes are next-generation DNA-binding dyes that are ideal for quantitative real-time PCR (qPCR) and many other applications. Biotium scientists designed the dyes by taking into consideration several essential dye properties relevant to PCR, including PCR inhibition, safety, stability and fluorescence spectra of the dye. The result of our efforts is a dye superior to SYBR® Green I and other commercial PCR or high-resolution melt curve (HRM) dyes.*

EvaGreen® Dye is used in all of Biotium’s dye-based qPCR master mixes, which combine superior brightness and sensitivity, with the ability to do sensitive melt curve analysis in the same reaction. EvaGreen® Dye is non-toxic, non-mutagenic, and not hazardous to aquatic life.

EvaGreen® Plus Dye is an improved alternative to Biotium’s original EvaGreen® Dye which retains the same essential benefits while providing a greater signal-to-noise ratio. The lower background and higher signal from EvaGreen® Plus provides further advantages for digital PCR and isothermal applications.

See EvaGreen® Dye Technology to learn more.

* Practicing HRM may require a license from Idaho Technologies, Inc.

Figure 1. EvaGreen® dye binds to dsDNA via a
Figure 1. EvaGreen® Dye binds to dsDNA via a “release-on-demand” mechanism.
Figure 2. EvaGreen® is non-toxic, non mutagenic and non hazardous to aquatic life.
Product NameCatalog NumberSize
EvaGreen® Dye, 20X in Water (trial size)31000-T1 mL
EvaGreen® Dye, 20X in Water 310005 x 1 mL
EvaGreen® Dye, 2000X in DMSO3101950 uL
EvaGreen® Plus Dye, 20X in Water (trial size)31077-T1 mL
EvaGreen® Plus Dye, 20X in Water310775 x 1 mL

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EvaGreen® is the only DNA-binding dye validated for droplet digital PCR (ddPCR)

See the full list of references here.

Highlighted Citation: The superior performance demonstrated by EvaGreen® dye has allowed it to be the first and only DNA-binding dye suitable for ddPCR applications. Initially designed for fluorescent probe-based detection, ddPCR technology has enabled quantitative multiplex PCR by partitioning samples into thousands of droplets that can be individually measured. In a publication in Analytical Chemistry, McDermott et al. demonstrate how EvaGreen® dye can be used with ddPCR to analyze multiple targets within a sample by distinguishing the amplified DNA mass per droplet. Results show EvaGreen®-based ddPCR performs comparably with TaqMan®-based ddPCR, without the added complexity and cost of fluorescent probes. EvaGreen® Dye is currently licensed for Bio-Rad’s QX200 ddPCR™ EvaGreen® Supermix and other ddPCR related reagents.

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Forget-Me-Not™ qPCR Master Mix is a hot-start EvaGreen® Dye-based master mix for use in real time PCR applications and DNA melt curve analysis. Forget-Me-Not™ features a unique combination of a master mix containing a low concentration of blue dye, plus a DNA template buffer containing a higher concentration of blue dye. When you add the 2X Forget-Me-Not™ Master Mix to your reaction, it appears light blue. Then, when you add template containing Forget-Me-Not™  Template Buffer to the reaction, the color turns dark blue (Figure 1, right). Forget-Me-Not™ allows you to see at a glance whether you forgot to add master mix or template to any of your reactions, so you can catch pipetting mistakes and avoid wasting time, reagents, and your precious DNA samples.

To learn more about our master mixes, please see Forget-Me-Not™ Master Mix Technology.

evagreen vs. sybr green
Figure 3. A comparison of the raw fluorescence signal from qPCR reactions performed with two EvaGreen® master mixes (Forget-Me-Not™ EvaGreen® and Fast EvaGreen®) and QuantiNova SYBR® Green. EvaGreen® dye is less inhibitory than SYBR® green, allowing for a much brighter signal.


Forget-Me-Not™ Universal Probe qPCR Master Mix is a high-performance product for fluorescent probe-based PCR applications, including quantitation and SNP genotyping. This kit is suitable for all fluorescent probe-based technologies, including hydrolysis probes (such as TaqMan® and dual-labeled BHQ® probes) and displacement probes (like Molecular Beacons). Forget-Me-Not™ Universal Probe Master Mix shows excellent concordance of results in singleplex and multiplex reactions, has broad instrument compatibility, and can be used in both standard and fast protocols. A separate kit is also available with ROX, if your instrument requires ROX reference dye.

To learn more about our master mixes, please see Forget-Me-Not™ Master Mix Technology.

forget me not universal probe master mix
Figure 4. Forget-Me-Not Universal Probe qPCR Master Mix works exceptionally well for mutiplexed probe PCR reactions.

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New: N-FLUX™ 5X Digital PCR Master Mix

Introducing our new N-Flux™ 5X Digital PCR Master Mix, a high-performance, dye-free master mix specially formulated for chip-based digital PCR (cdPCR) platforms. The master mix is designed to minimize interactions with hydrophobic plastic surfaces for optimal sample partitioning, resulting in maximum end-point fluorescence and superior separation between positive and negative partitions. N-Flux™ 5X Digital PCR Master Mix has been validated in singleplex and multiplex probe-dPCR reactions. Using the Combinati Absolute Q digital PCR platform, N-Flux™ 5X digital PCR Master Mix reproducibly performed genotyping and copy number variation analysis with quantitative discrimination. In addition, the high 5X concentration simplifies workflows for users performing qPCR on large sample volumes. This offers flexibility and facilitates PCR analysis for samples that are too dilute for traditional 2X master mixes.

To learn more, please see our N-Flux™ 5X Digital PCR Master Mix product page.


  • Robust results:
    Sensitive and reproducible performance in dPCR and qPCR assays.
  • Optimal sample partitioning:
    Minimizes interactions with hydrophobic plastic surfaces and maximizes end-point fluorescence.
  • Compatible with all fluorescent probe-based technologies:
    Including TaqMan® dual-labeled such as BHQ® probes, and displacement probes.
  • Convenient 5X concentration for large sample volumes
    Facilitates workflows for samples that are too dilute for 2X master mixes.
High-resolution, multiplex copy number variation analysis performed using N-Flux™ 5X Digital PCR Master Mix on the COMBiNATi Microfluidic Array Partitioning (MAP16) device. Superior partitioning results in quantitative determination of the copy number of a low abundance SNP variant and a high abundance control on human gDNA, with excellent separation between positive and negative wells.
Product NameCatalog NumberSize
N-Flux™ 5X Digital PCR Master Mix (trial size)31078-T50 x 20 uL reactions
N-Flux™ 5X Digital PCR Master Mix31078250 x 20 uL reactions

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Master Mix Selection Guide

Catalog No.Master MixEvaGreen® qPCRProbe-Based PCROptimized for dPCR1-color Tracking2-Color Tracking
31041Forget-Me-Not™ EvaGreen® qPCR Master Mix with 2-Color Tracking
31042Forget-Me-Not™ EvaGreen® qPCR Master Mix with 2-Color Tracking,
with separate ROX
31045Forget-Me-Not™ EvaGreen® qPCR Master Mix (low ROX)
31046Forget-Me-Not™ EvaGreen® qPCR Master Mix (high ROX)
31043Forget-Me-Not™ Universal Probe qPCR Master Mix (no ROX)✓*
31044Forget-Me-Not™ Universal Probe qPCR Master Mix, with separate ROX✓*
31078N-Flux™ 5X Digital PCR Master Mix


Cheetah™ Taq is a chemically-modified hot-start DNA polymerase. Using a proprietary method developed at Biotium, Cheetah™ Taq is modified to have absolutely no activity at room temperature, but be completely activated after two minutes at 95°C.


  • Fast activation:
    Takes less than 2 minutes to activate, much faster than AmpliTaq Gold and HotStarTaq®.
  • Excellent activity recovery:
    Better recoveries of both 5′-3′-polymerase and 5′-exonuclease activities following activation than AmpliTaq Gold.
  • Improved stability:
    Better stability than AmpliTaq Gold during storage.
  • pH compatibility:
    Can be activated at alkaline pH (8.5-9) for optimal Taq activity and more specific target amplification.
hot start enzymes
Figure 5. Cheetah™ Taq recovers full activity faster than other hot-start enzymes such as AmpliTaq Gold.
Product NameCatalog NumberSize
Cheetah™ HotStart Taq DNA Polymerase29050500 U

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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. In the presence of DNA polymerase activity, the primer will be extended to form a double stranded product that can bind EvaGreen® Dye, resulting in an increase of fluorescence. The rate of increase of fluorescence is positively correlated to the activity of polymerase. The assay was developed for the measurement of Taq DNA polymerase activity. It can also be used for other DNA polymerases such as Pfu, Vent, Phusion®, Bst, Phi29, MMLV, AMV, SuperScript®, T4 DNA polymerase, T7 DNA polymerase, Klenow, and E. coli DNA polymerase I. The activity assay can be conducted at temperatures from 4°C to 75°C.

evaez assays taq dna polymerase
Figure 6. Sample data from an EvaEZ assays, showing a titration of Taq DNA Polymerase.
Product NameCatalog NumberSize
EvaEZ™ Fluorometric Polymerase Activity Assay Kit29051200 assays

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Biotium is proud to have developed the cutting-edge technology of viability PCR, a technique in which cell viability can be assessed by treating a sample with a DNA-binding dye, followed by a qPCR or isothermal amplification reaction. To learn more about this technology and our portfolio of reagents for viability PCR, please see Viability PCR Technology.

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Biotium offers a wide range of PCR/qPCR accessory products to assist you in your DNA amplification experiments, including:

  • dNTPs, sold as mixes or in separate tubes
  • Fluorescently-labeled and biotinylated nucleotides (dUTP, dCTP & others)
  • ROX Reference Dye
  • PCR dye (see EvaGreen®)
  • Taq DNA Polymerase (see Cheetah™ Taq)
  • HotStart and WarmStart™ Polymerase Modification Kits
Product NameCatalog NumberSize
dNTP Mix, 10 mM Each400545 x 1 mL
dNTP Mix, 25 mM Each400531 mL
dNTP Set, 100 mM Each40052250 uL Each
5-Aminoallyl dUTP (AA-dUTP), 10 mM Solution40020100 uL
BrdU40024100 mg
dCTP CF® Dye Conjugates9 dye color options5 or 25 nmol
dUTP CF® Dye Conjugates8 dye color options5 or 25 nmol
ROX Reference Dye290525 x 1 mL
EvaGreen® Dye, 2000X in DMSO3101950 uL
EvaGreen® Dye, 20X in water310005 x 1 mL
Cheetah™ Hot-Start Taq Polymerase29050500 U
HotStart Polymerase Modification Kit290541 kit modifies 0.5 mg of polymerase
WarmStart™ Enzyme Modification Kit290531 kit modifies 0.5 mg of protein

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

From: $63 Sizes: 1 mL (100 reactions) Trial Size, 5 x 1 mL (500 reactions), 2 x 10 mL (2000 reactions)Catalog #: 31041-T, 31041-1, 31041-20mL, 31042-T, 31042-1, - 31042-20mLView allHide

Forget-Me-Not™ Universal Probe qPCR Master Mix

From: $63 Sizes: 1 mL (100 reactions) Trial Size, 5 x 1 mL (500 reactions)Catalog #: 31043-1, 31043-T, 31044-1, - 31044-TView allHide

EvaGreen® Dye, 20X in Water

From: $47 Sizes: Trial size (1 mL), 5 x 1 mLCatalog #: 31000, - 31000-TView allHide

Cheetah™ HotStart Taq DNA Polymerase

From: $239 Sizes: 500 UCatalog #: 29050

EvaEZ™ Fluorometric Polymerase Activity Assay Kit

From: $398 Sizes: 2 x 1 mLCatalog #: 29051

HotStart Polymerase Modification Kit

From: $210 Sizes: Trial size, 1 kitCatalog #: 29054, - 29054-TView allHide

WarmStart Enzyme Modification Kit

From: $210 Sizes: Trial size, 1 kitCatalog #: 29053, - 29053-TView allHide

dNTP Mix, 10 mM Each

From: $397 Sizes: 5 x 1 mLCatalog #: 40054