EvaGreen® qPCR Supermix

Suitable for ROX-dependent and ROX-independent qPCR cyclers

For in vitro use only

PACK SIZES : 1ml | 8ml | 20ml

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HOT FIREPol® EvaGreen® qPCR Supermix is an optimised ready-to-use solution for real-time quantitative PCR assays, including EvaGreen® dye. It comprises all the components necessary to perform qPCR excluding the template and primers, to perform highly sensitive qPCR.
HOT FIREPol®DNA Polymerase is activated by a 12 min incubation step at 95°C. Hot-start mechanism prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.

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– Detection and quantification of DNA and cDNA targets

– Profiling gene expression

– Microbial detection

– Viral load determination



– Highly specific and reproducible real time PCR

– Excellent efficiency in case of low copy number targets

– UNG treatment capability due to dNTP blend of dUTP/dTTP

– Superior performance with long (up to 500 bp) and GC-rich templates

– Blue visualisation dye for easy pipetting

Mix Compatibility:

– Appilied BioSystems: QuantStudio™ 12K Flex, ViiA™ 7, 7900HT, 7500, 7700, StepOneTM & StepOnePlusTM

– Stratagene: MX3000PTM , MX3005PTM

Bio-Rad: CFX96™ & CFX384™, iQ™5 & MyiQ™, Chromo4™, Opticon® 2 & MiniOpticon®

Qiagen: Rotor-Gene® Q, Rotor-Gene® 6000 

Eppendorf: Mastercycler®: ep realplex2 & ep realplex4 

llumina: The Eco™  

Roche: LightCycler® 480


Mix Composition:

– HOT FIREPol® DNA Polymerase

– 5x EvaGreen® qPCR buffer

– 12.5 mM MgCl2 1x PCR solution – 2.5 mM

– dNTPs blend containing dUTP/dTTP including dTTP to improve reaction sensitivity and efficiency compared to dUTP 

– EvaGreen® dye 

– Internal reference based on ROX dye

– GC-rich Enhancer

– Blue visualisation dye


EvaGreen® Dye:

EvaGreen® is a DNA-binding dye with many features that make it a superior alternative to SYBR® Green I for qPCR. Apart from having similar spectra, EvaGreen® has three important features that set it apart from SYBR® Green I: EvaGreen® has much less PCR inhibition, is extremely stable dye and has been shown to be nonmutagenic and noncytotoxic. EvaGreen® is compatible with all common real-time PCR cyclers – simply select the standard settings for SYBR® Green or FAM!


ROX Dye:

ROX is an internal passive reference dye used to normalize the fluorescent reporter signal generated in qPCR.

Shipping and Storage Conditions:

Routine storage: -20ºC
Shipping and temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of HOT FIREPol® EvaGreen® qPCR Supermix.


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