HOT FIREPol® EvaGreen® qPCR Supermix

HOT FIREPol^® EvaGreen^® qPCR Supermix is an optimized 5X reaction mix for dye-based qPCR detection and quantitation of target DNA sequences using EvaGreen^® dye and SYBR^®/FAM channel of most real-time qPCR instruments.

The Mix contains HOT FIREPol^® DNA Polymerase and a passive internal reference dye that is compatible with most qPCR cyclers including no ROX and low ROX* reference signal requiring platforms. 

The Mix contains dUTPs to prevent cross-contamination with UNG treatment and visible dye to ease reaction set-up. This dye does not interfere with real-time detection.

Applications

• Detection and quantification of DNA and cDNA targets
• Profiling gene expression
• Microbial detection
• Viral load determination

Properties

Concentration: 5x

Hot-start: yes, initial activation in 12 min

Detection type: dye-based, includes EvaGreen^® intercalating dye

Reference dye*: based on ROX

Compatible real-time instruments: most qPCR cyclers. Check Your cycler!

IMPORTANT UPDATE! HOT FIREPol^® EvaGreen^® qPCR Supermix is not compatible with high ROX cyclers such as Applied BioSystems^® StepOne™ or StepOnePlus™

Mix Components

HOT FIREPol^® DNA polymerase: chemically modified FIREPol^® DNA Polymerase enabling hot-start

5x EvaGreen^® qPCR buffer with 12. 5 mM MgCl₂: 1x PCR solution – 2.5 mM MgCl₂

dNTPs, including dUTP: mix allows UNG treatment to prevent carryover contamination from previous runs.
IMPORTANT: UNG is not included in the HOT FIREPol^® EvaGreen^® Supermix

EvaGreen^® dye

Internal reference based on ROX dye

GC-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 an extremely stable dye, and has been shown to be non-mutagenic and non-cytotoxic. EvaGreen^® is compatible with all common real-time PCR cyclers – simply select the standard settings for SYBR^® Green or FAM!