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Bright, sensitive and environmentally safe; ideal for qPCR and many other applications
A spectrally unique red qPCR dye that expands probe-based multiplexing
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.
Chemically modified hot-start taq; no activity at RT and activates in only 2 minutes
Offers reliable and steady PCR normalization. Functions as ROX alternative and direct replacement for Mustang Purple™.
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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EvaRuby™ Dye for qPCR & HRM® Analysis

A spectrally unique intercalating dye that expands multiplexing options for probe-based qPCR

EvaRuby™ Dye is the first visible red DNA-binding dye for both qPCR and HRM. The dye has a long Stokes shift of 480/613 nm (excitation/emission), allowing excitation  by the same green/blue channel as EvaGreen® Dye, and detection in the orange (JUN™ or ROX™) channel. As a result, EvaRuby™ Dye has minimal cross-talk in standard probe detection channels and enables the combination of dye-based and probe-based qPCR in a single reaction. In addition, EvaRuby™ Dye is validated for HRM® analysis and may be used as an internal control for probe-based qPCR to detect non-specific amplification that goes undetected by target-specific probes. EvaRuby™ Dye is also suitable for singleplex qPCR, LAMP, and gel electrophoresis.

  • Flexible: Unique orange/red DNA-binding dye that can be combined with qPCR probes to expand multiplexing capabilities
  • Versatile: Can combine probe-based qPCR with HRM® in the same reaction
  • Spectrally Unique: Minimal cross-talk in neighboring channels
  • Convenient: Easily confirm your results with melt curve analysis rather than time consuming post-qPCR gel electrophoresis
EvaRuby™ Dye, 20X in Water
High Resolution Melt Data Difference Plot. EvaRuby™ Dye at 1X in Forget-Me-Not™ Universal Probe qPCR Master Mix (Cat. No. 31043) was used to detect the Applied Biosystems® MeltDoctor™ HRM Positive Control Kit. Data was acquired using the Qiagen® Rotor-Gene® Q, 0.1°C/2 sec, with a 470 nm excitation and 610 nm emission filter. Data analysis was performed with uAnalyzeSM ( using baseline subtraction. EvaRuby™ Dye can clearly distinguish each homozygous variant as well as the heterozygous variant.

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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 (Fig 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.

Figure 5. 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.

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Full List of Master Mixes

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.
Cheetah™ HotStart Taq DNA Polymerase – 500 U
hot start enzymes
Figure 6. Cheetah™ Taq recovers full activity faster than other hot-start enzymes such as AmpliTaq Gold.

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VeriFluor™ Far-Red Passive Reference Dye

VeriFluor™ Far-Red is a far-red passive reference dye that offers reliable and steady fluorescent signal for normalizing the reporter dye during PCR amplification. VeriFluor™ acts as a direct replacement for Mustang Purple™ and as an alternative to ROX passive reference dye, allowing more flexibility when designing multiplex qPCR reactions. Unlike Mustang Purple™, VeriFluor™ also has demonstrated insensitivity to common buffer additives such as DTT (Fig 7, right).

  • Offers reliable and steady fluorescent signal for normalizing qPCR reactions
  • An alternative to ROX, allowing more flexibility for multiplexing
  • Drop-in replacement for Mustang Purple™ on most instruments
  • Tolerates DTT in qPCR reactions, unlike Mustang Purple™
  • More soluble than ROX for easier mixing and more accurate results
  • Stand-alone dye solution for use with our Forget-Me-Not™ Master Mixes or other mixes
VeriFluor™ Far-Red Passive Reference Dye, 400X in Water – 50 uL
VeriFluor™ Far-Red Passive Reference Dye, 400X in Water – 500 uL
Figure 7. Non-normalized qPCR plot with Forget-Me-Not™ Master Mix with EvaGreen® (Greens) and VeriFluor™ Far-Red Passive Reference (Purples) on the QuantStudio 5 qPCR instrument. There is no distinguishable difference between VeriFluor™ Far-Red or EvaGreen® signal with or without 4mM DTT.

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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™ Fluorometric Polymerase Activity Assay Kit – 2 x 1 mL
evaez assays taq dna polymerase
Figure 6. Sample data from an EvaEZ assays, showing a titration of Taq DNA Polymerase.

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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: $73 Sizes: 1 mL (100 reactions) Trial Size, 5 x 1 mL (500 reactions), 2 x 10 mL (2000 reactions)Catalog #:
, 31041-1, 31041-20mL, 31042-T, 31042-1, - 31042-20mLView allHide

Forget-Me-Not™ Universal Probe qPCR Master Mix

From: $73 Sizes: 1 mL (100 reactions) Trial Size, 5 x 1 mL (500 reactions)Catalog #:
, 31043-T, 31044-1, - 31044-TView allHide

EvaGreen® Dye, 20X in Water

From: $54 Sizes: Trial size (1 mL), 5 x 1 mLCatalog #:
, - 31000-TView allHide

Cheetah™ HotStart Taq DNA Polymerase

From: $277 Sizes: 500 UCatalog #: 29050

EvaEZ™ Fluorometric Polymerase Activity Assay Kit

From: $498 Sizes: 2 x 1 mLCatalog #: 29051

HotStart Polymerase Modification Kit

From: $242 Sizes: Trial size, 1 kitCatalog #:

WarmStart™ Enzyme Modification Kit

From: $242 Sizes: Trial size, 1 kitCatalog #:

dNTP Mix, 10 mM Each

From: $460 Sizes: 5 x 1 mLCatalog #: 40054

Ultrafiltration Vials

From: $53 Sizes: 5 itemsCatalog #:
, - 22018View allHide

EvaRuby™ Dye, 20X in Water

From: $99 Sizes: 500 uL, 2 x 1 mLCatalog #:
, - 31079-TView allHide

VeriFluor™ Far-Red Passive Reference Dye, 400X in Water

From: $111 Sizes: 50 uL, 500 uLCatalog #:
, - 29087-500uLView allHide

N-Flux™ 5X Digital PCR Master Mix

From: $99 Sizes: Trial size (1 x 200uL), 5 x 200uLCatalog #:
, - 31078View allHide