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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. 

Sensitive qPCR Mix designed for effective quantification of DNA targets with probe-based chemistry

Cost-effective real time qPCR master mix for probe-based qPCR assays.
This master mix has been developed for TaqMan® probes but is suitable for other hydrolysis probe types as well.

  • suitable for singleplex and duplex assays
  • high specificity and sensitivity
  • reaction set-up and shipment without dry ice

Ordering

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Catalog number

08-16-0000S

Quantity

€ 0.00

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Details

Overview

Description

HOT FIREPol® Probe qPCR Mix Plus (Capillary) is optimized for real-time quantitative PCR assays and contains all the components necessary to perform qPCR, with the exception of template, primers, and probe.

The qPCR Mix contains optimized components and HOT FIREPol® DNA Polymerase supplied in a proprietary reaction buffer that enables detection of low copy number targets. HOT FIREPol® Probe qPCR Mix Plus (Capillary) is optimized for DNA hydrolysis probes based on the 5´ flap endonuclease activity.

HOT FIREPol® DNA Polymerase is activated by a 12 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 12-15 min

Detection type: Probe-based

Reference dye: none

Suitable qPCR Cyclers: LightCycler 1.x and 2.0 (Roche Applied Science). Check Your cycler!

Mix Components

HOT FIREPol® DNA Polymerase: chemically modified FIREPol® DNA Polymerase enabeling hot-start

5x Probe qPCR buffer with 15 mM MgCl2: 1x PCR solution – 3 mM MgCl2

dNTPs: dATP, dCTP, dGTP and dTTP

Bovine serum albumine (BSA) 

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.

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.

Publications

Detection of HEV in naturally infected swine from central Argentina by an optimized HEV/MS2 duplex RT-qPCR

F. Marziali et al. / Zoonoses Public Health / 2019

Which sympathoadrenal abnormalities of adult spontaneously hypertensive rats can be traced to a prehypertensive stage?

A. Vavřínová et al. / Hypertens. Res. / 2019

Alterations to the adhesion, growth and osteogenic differentiation of human osteoblast-like cells on nanofibrous polylactide scaffolds with diamond nanoparticles

L. Stankova et al. / Diam. Relat. Mater. / 2019
read more

miR-10a-5p is increased in atopic dermatitis and has capacity to inhibit keratinocyte proliferation

H. Vaher et al. / Allergy Eur. J. Allergy Clin. Immunol. / 2019

Orexin/hypocretin receptor gene (HCRTR1) variation is associated with aggressive behaviour

J. Harro et al. / Neuropharmacology / 2019

Antioxidant, Anti-Inflammatory, and Multidrug Resistance Modulation Activity of Silychristin Derivatives

J. Viktorová et al. / Antioxidants / 2019

Detection of Murine Astrovirus and Myocoptes musculinus in individually ventilated caging systems: Investigations to expose suitable detection methods for routine hygienic monitoring

C. Körner et al. / PLoS One / 2019

Dexamethasone resets the circadian clock in hippocampus via multiple mechanisms involving lithium‐independent GSK3β signalling

K. Šuchmanová et al. / Br. J. Pharmacol. / 2019

SERPINB2 and miR‐146a/b are coordinately regulated and act in the suppression of psoriasis‐associated inflammatory responses in keratinocytes

H. Vaher et al. / Exp. Dermatol. / 2019

Microbiota affects the expression of genes involved in HPA axis regulation and local metabolism of glucocorticoids in chronic psychosocial stress

M. Vodička et al. / Brain. Behav. Immun. / 2018

Telomere length and survival in primary cutaneous melanoma patients

S. Rachakonda et al. / Sci. Rep. / 2018

Lipopolysaccharide- and β-1,3-glucan-binding protein from Litopenaeus vannamei: Purification, cloning and contribution in shrimp defense immunity via phenoloxidase activation

B. Phupet et al. / Dev. Comp. Immunol. / 2018

Utilization of organic nitrogen by arbuscular mycorrhizal fungi—is there a specific role for protists and ammonia oxidizers?

P. Bukovská et al. / Mycorrhiza / 2018

FSHB -211 G>T is a major genetic modulator of reproductive physiology and health in childbearing age women

K. Rull et al. / Hum. Reprod. / 2018
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® Probe qPCR Mix Plus.

Shipping

At room temperature