Some applications of this product may require a license which is not provided by the purchase of this product.
For research use only.
Previous version of this product, HOT FIREPol® SolisGreen® qPCR Mix (cat. no 08-46-XXXXX) has been discontinued. HOT FIREPol® SolisGreen® qPCR Mix 2.0 shows identical performance as per product specification with its previously existing product version.
HOT FIREPol® SolisGreen® qPCR Mix 2.0 is a 5x-concentrated solution specially designed for dye-based real-time quantitative PCR (qPCR) assays. SolisGreen® dsDNA binding dye is incorporated into the product to evaluate the DNA amplification process during PCR. The fluorescence spectra of this dye are similar to the more widely used SYBR® Green I and is compatible with most major real-time cyclers. When compared to SYBR® Green I, SolisGreen® dye shows:
HOT FIREPol® SolisGreen® qPCR Mix 2.0 is an optimized ready-to-use solution that contains all the necessary components, except sample (DNA template) and primers, to perform reactions with accurate and highly sensitive results. Additionally, the product includes a passive reference based on ROX dye making it compatible with both ROX-dependent and ROX-independent qPCR cyclers.
The product contains HOT FIREPol® DNA Polymerase to prevent the extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.
For RNA applications try out an already optimized easy-to-use SOLIScript® 1-step SolisGreen® Kit 2.0 or combine HOT FIREPol® SolisGreen® qPCR Mix 2.0 with thermostable reverse transcriptases, such as FIREScript® and SOLIScript®!
Sample type: DNA
Concentration: 5x
Hot-start: Yes, initial activation in 10 min
Detection type: Dye-based, includes SolisGreen® intercalating dye
Reference dye: Based on ROX
Compatible real-time instruments: The mix is compatible with both ROX-dependent and ROX-independent qPCR cyclers. HOT FIREPol® SolisGreen qPCR Mix 2.0 is not compatible with platforms that require high ROX, such as Applied BioSystems® 7900HT, StepOne™ or StepOnePlus™ systems. Check Your cycler!
HOT FIREPol® DNA polymerase: chemically modified FIREPol® DNA Polymerase enabling hot-start.
5x qPCR buffer with 12.5 mM MgCl2: 1x PCR solution – 2.5 mM MgCl2
dNTPs: dATP, dCTP, dGTP and dTTP
SolisGreen® dye
Internal reference based on ROX dye
Dye-based detection (e.g., EvaGreen®, SolisGreen®, SYBR® Green) is a cost-effective qPCR option, as it requires only addition of PCR primers. However, the intercalating dye will detect any dsDNA (non-specific amplicons, primer dimers) produced in the reaction. Melt curve analysis performed after the qPCR can be used to verify the specificity of amplification and to check for the presence of non-specific amplification products.
The probe-based qPCR system demonstrates higher specificity compared to dye-based qPCR, because probes only detect the gene of interest. Keep in mind that in probe-based assays, primer dimers and non-specific products will not be detected, however, they may compromise the PCR efficiency. Using probe-based qPCR system, it is possible to distinguish between sequences with high similarity (e.g. single-nucleotide variations). Additionally, probe-based qPCR assays allow for multiplex reactions in one tube, while only a single target can be amplified and measured in a dye-based qPCR.
SolisGreen™ nucleic acid dye shares spectral properties with other commonly used qPCR dyes, allowing detection in the FAM or SYBR® Green I channel (SolisGreen™ excitation maximum 489 nm, emission maximum 515 nm). SolisGreen™ is characterized by high sensitivity and great PCR efficiency with low template amounts for accurate and reproducible results.
SolisGreen™ is based on CYGREEN dye. CYGREEN is used under licence from Enzo Life Sciences, Inc. CYGREEN is a U.S. registered trademark of Enzo Life Sciences, Inc. U.S. Patent Nos. 8,153,802 and 7,569,695.
Both dyes are DNA intercalating agents that are used to stain dsDNA. SolisGreen™ dye is spectrally similar to EvaGreen® allowing detection in the FAM or SYBR® Green I channel (SolisGreen™ excitation maximum 489 nm, emission maximum 515 nm). It has been shown that qPCR mixes based on SolisGreen™ dye have increased performance with low target concentration demonstrating improved precision and less variance between technical replicates.
Recommended final amount of cDNA sample in downstream PCR reaction is up to one tenth of the final reaction volume. Overload of cDNA sample may compromise the downstream PCR, because cDNA sample may contain reaction components that may inhibit your PCR reaction.
ROX (carboxy-X-rhodamine) is one of the passive reference dyes providing a constant fluorescent signal that is used for signal normalization during the amplification cycles. The emission recorded from a reference dye during the baseline cycles is used to normalize the emission recorded from the reporter (e.g. EvaGreen®, SolisGreen®, SYBR® Green, etc.) in later cycles in ROX-dependent real-time PCR systems (e.g. Applied Biosystems 5700, 7000, 7300, 7700, 7900HT, StepOne™, StepOnePlus™). Reference dye compensates for small fluorescent fluctuations and well-to-well variations that may occur.
In Solis BioDyne qPCR mixes we use technology, based on ROX, that allows us to use same mix for high- and low- ROX requiering cyclers.
Please check cycler-mix compatibility here if not sure.
Solis BioDyne products should be stored at -20°C.
Shipping and temporary storage for up to 1 month at room temperature (*15−25°C) has no detrimental effects on the quality of Solis BioDyne reagents.
Freeze-thaw stability is tested for each product. Most PCR and qPCR products have passed 30 freeze-thaw cycles with no changes in performance. Specific information is found in Storage and Shipping conditions of each product.
When stored and handled under the recommended conditions, full activity of the reagents is retained until the Expiry Date printed on the tube label.
*World Health Organization (2003). Guidelines for the Storage of Essential Medicines and Other Health Commodities.
Routine storage: -20ºCTemporary storage for up to 1 month at room temperature has no detrimental effects on the quality of HOT FIREPol® SolisGreen® qPCR Mix 2.0.
At room temperature.
Ready-to -use solution for simple dye-based qPCR assays, incorporating EvaGreen® dye. Available for ROX, no ROX, and capillary cyclers.
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Extremely fast and highly sensitive dye-based qPCR that includes novel in-silico designed SolisFAST® DNA Polymerase Mix for most demanding applications
Highly sensitive cDNA synthesis and dye-based qPCR in a single reaction. Ideal solution for cost effective and high throughput RNA analysis.
A flexible and robust cDNA synthesis kit contains all the reagents you need for first-strand cDNA synthesis. Individual components enable thorough assay optimization.
A flexible and highly specific cDNA synthesis kit that contains all the reagents you need for first-strand cDNA synthesis. Individual components enable thorough assay optimization.
