MBI EVOlution 5 x EvaGreen® qPCR Mix (no 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: MBI Hotstart DNA Polymerase, ultrapure dNTPs, MgCl2 and EvaGreen® dye. The user simply needs to add water, template and primers.
MBI Hotstart DNA Polymerase is activated by a 15 min incubation step at 95°C. This prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.
Applications
Detection and quantification of DNA and cDNA targets
Profiling gene expression
High Resolution Melt (HRM) Analysis
Microbial detection
Viral load determination
Mix Composition
MBI Hotstart DNA Polymerase
5 x EvaGreen® qPCR buffer
12.5 mM MgCl2 (1x PCR solution = 2.5 mM MgCl2)
dNTPs, including dTTP to improve reaction sensitivity and efficiency compared to dUTP
EvaGreen® dye
No 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 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!
Recommended qPCR Reaction Mix
Component
Volume
Final conc.
MBI EVOlution 5 x EvaGreen® qPCR Mix (no ROX)
4 µl
1x
Primer Forward (10 pmol/µl)
0.16-0.5 µl
80-250 nM
Primer Reverse (10 pmol/µl)
0.16-0.5 µl
80-250 nM
DNA template
1-5 µl
1-50 ng/µl
H2O PCR grade
up to 20 µl
-
Total
20 µl
-
Recommended qPCR Cycles
Cycle step
Temp.
Time
Cycles
Initial denaturation
95ºC
15 min
1
Denaturation
95ºC
15 s
40
Annealing
60º-65ºC
20 s
Elongation
72ºC
20 s
IMPORTANT: To activate the polymerase, include an incubation step at 95ºC for 15 minutes at the beginning of the qPCR cycle.
Safety Warnings and Precautions
This product and its components should be handled only by persons trained in laboratory techniques. It is advisable to wear suitable protective clothing, such as laboratory overalls, gloves and safety glasses. Care should be taken to avoid contact with skin or eyes. In case of contact with skin or eyes, wash immediately with water.
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 MBI EVOlution 5 x EvaGreen® qPCR Mix (no ROX).
EvaGreen® is a registered trademark of BIOTIUM, INC.
The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer, where such research does not include testing, analysis or screening services for any third party in return for compensation on a per test basis. The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes.
SYBR® is a registered trademark of Molecular Probes, Inc.
Some applications this product is used in may require a license which is not provided by the purchase of this product. Users should obtain the license if required.
Montreal Biotech Inc. Quality Laboratory Equipment for Research & Industry www.montreal-biotech.com
