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.

See the full list of references here.

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

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.

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.

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.

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.

FEATURES

• 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.

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

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.