5x HOT FIREPol EvaGreen® qPCR Mix Plus (ROX)

Supplier
LabOmics Solis Biodyne
Cat. No
6330250
Pack size
1ml (250rxn)
Links
Data Sheet MSDS

Features

• much less PCR inhibition compare to SYBR® Green
• extremely stable dye
• nonmutagenic and noncytotoxic

HOT FIREPol® EvaGreen® qPCR Mix Plus(ROX) is an optimised ready-to-use solution for real-time quantitative PCR assays, incorporating EvaGreen® dye. It comprises all the components necessary to perform qPCR: HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2, EvaGreen® dye and ROX dye according to system requirements. The user simply needs to add water, template and primers. HOT FIREPol® DNA Polymerase is activated by a 15 min incubation step at 95°C. This prevents extension of nonspecifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.

• 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. The high ROX concentration in the final 1x reaction is 500 nM.
• Cycler Compatibility: HOT FIREPol® EvaGreen® qPCR Mix (high ROX) is compatible with all qPCR cyclers requiring high ROX dye level, including ABI PRISM® 7000, 7300, 7700, 7900 and 7900HT (including Fast-Block).
• 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 Mix (high ROX).
• Mix Composition:

• HOT FIREPol® DNA Polymerase
• 5 x EvaGreen® qPCR buffer
• 15 mM MgCl2

1 x PCR solution – 3 mM MgCl2

• dNTPs, including dTTP to improve reaction sensitivity and efficiency compared to dUTP
• EvaGreen® dye
• 2.5 µM ROX dye

1 x PCR solution – 500 nM ROX dye

Log scale amplification plot of FBA1 gene.

Amplification of a 92 bp fragment from FBA1 gene was performed on serial fourfold dilutions of Saccharomyces cerevisiae genomic DNA. Reactions were performed on an Applied Biosystems 7900HT Real-Time PCR System and ROX was used as a passive reference for normalisation of the reporter signal.

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