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Some applications of this product  may require a license which is not provided by the purchase of this product.

For research use only. 

Highly sensitive dye-based qPCR Mix with improved detection of low-copy targets.

Optimized ready-to-use solution for dye-based real-time quantitative
PCR assays. SolisGreen® dye is characterized by high sensitivity and
great PCR efficiency with low template amounts for accurate and
reproducible results

  • improved detection of low target concentrations
  • higher fluorescence level
  • reaction set-up and shipment without dry ice

Ordering

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Catalog number

08-46-00001

Quantity

€ 72.00

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Details

Overview

Description

Dye-based real-time quantitative PCR (qPCR) uses DNA binding dye to evaluate DNA amplification process during PCR. 

HOT FIREPol® SolisGreen® qPCR Mix is an optimised ready-to-use solution for real-time quantitative PCR assays, incorporating SolisGreen® dye.

It comprises all the components necessary, excluding the template and primers, to perform highly sensitive qPCR. The user simply needs to add water, template and primers.

HOT FIREPol® DNA Polymerase is activated by a 10 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
  • Microbial detection
  • Viral load determination

Properties

Concentration: 5x

Hot-start: yes, initial activation in 10 min

Detection type: dye-based, includes EvaGreen® intercalating dye

Reference dye: based on ROX

Compatible real-time instruments: Most qPCR cyclers except platforms that require high ROX (HOT FIREPol® SolisGreen qPCR Mix is not compatible with the Applied BioSystems® 7900HT, StepOne™ or StepOnePlus™ systems.) Check Your cycler!

Mix Components

HOT FIREPol® DNA polymerase: chemically modified FIREPol® DNA Polymerase enabeling 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

Bovine serum albumine (BSA) to enhance PCR reaction

Internal reference based on ROX dye

SolisGreen® dye

This dye-based qPCR mix contains next generation DNA binding dye  SolisGreen®. The fluorescence spectra of this dye is similar to the more widely used SYBR® Green I and is compatible with most major realtime cyclers. SolisGreen® dye shows:

  • high fluorescence level
  • high sensitivity for detecting low template concentrations
  • high stability for room temperature storage
Highly competitive
Excellent sensitivity
Related Resources

FAQ

What is the difference between dye-based and probe-based systems?

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.

I have previously used SYBR® Green / EvaGreen® based qPCR assays. What are the spectral features of SolisGreen™ dye?

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.

What is the difference between EvaGreen® and SolisGreen® dye?

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.

After first-strand cDNA synthesis, how much cDNA template should I use for my PCR?

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.

Why do certain kits contain a ROX?

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.

What are the storage conditions and stability of Solis BioDyne reagents?

Solis BioDyne products should be stored at -20°C.

Shipping and temporary storage for up to 1 month at room temperature (*1525°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.

Storage & Shipping
Storage

Routine storage: -20ºC
Temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of HOT FIREPol® SolisGreen qPCR Mix.

Shipping

At room temperature