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NEW!

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

  • Great PCR efficiency – reliable and easy-to-analyze results
  • High sensitivity – great when working with low concentrated samples
  • Excellent fluorescence levels
  • Convenient reaction set-up at room temperature
  • Environmentally friendly – worldwide ice-free shipping

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.

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Catalog number

08-47-0000S

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Details

Overview

Description

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:

  • higher fluorescence level
  • better sensitivity for detecting low template concentrations
  • great stability for room temperature storage

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.

Applications

  • Gene expression analysis and absolute quantification
  • Pathogen detection and quantification
  • Low-copy gene detection
  • Cell-free DNA analysis

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®!

Properties

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!

Mix Components

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

Related Resources

Documents

    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 2.0.

    Shipping

    At room temperature.