Dye-based real-time quantitative PCR (qPCR) uses DNA binding dye to evaluate the DNA amplification process during PCR. In this mix EvaGreen® double-stranded DNA binding dye is used instead of the more widely used SYBR Green I that has similar fluorescence spectra. Compared to SYBR Green I dye EvaGreen® dye shows a higher fluorescence level, high sensitivity for detecting low template concentrations, and high stability at room temperature.
HOT FIREPol® EvaGreen® qPCR Mix Plus (ROX) is an optimized ready-to-use solution for dye-based real-time quantitative PCR assays on cyclers that require passive reference dye (including high ROX or low ROX reference signal requiring platforms).
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 12-15 min
Detection type: dye-based, includes EvaGreen® intercalating dye
Reference dye: based on ROX
Compatible real-time instruments: Cyclers that require ROX reference dye. Check Your cycler!
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: dATP, dCTP, dGTP and dTTP
EvaGreen® dye
Internal reference based on ROX 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!