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!
