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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. 

Optimized ready-to-use solution for High Resolution Melt (HRM) Analysis

  • highly sensitive mix for accurate SNP detection
  • sensitive EvaGreen® dye for reliable HRM performance
  • reaction set-up without dry ice

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Catalog number

08-33-0000S

Quantity

€ 0.00

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Details

Overview

Description

High Resolution Melt (HRM) analysis is fast and cost-effective method for post-PCR analysis to detect single nucleotid polymorphisms (SNP genotyping), genetic mutations and DNA sequence variations.

5x HOT FIREPol® EvaGreen® HRM Mix is an optimised ready-to-use solution for HRM analysis, incorporating EvaGreen® dye.

It comprises all the components necessary to perform qPCR and HRM Analysis: HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2, EvaGreen® dye and ROX dye according to system requirements. The user simply needs to add water, template and primers.

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.

Applications

  • High Resolution Melt (HRM)
  • SNP detection
  • dye-based qPCR

Properties

Concentration: 5x

Hot-start: yes, initial activation in 12 min

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

Reference dye: based on ROX

Compatible real-time instruments: Cyclers that require ROX reference dye (Applied BioSystems). Check Your cycler!

Mix Components

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

5x EvaGreen® qPCR buffer with 12. 5 mM MgCl2: 1x PCR solution – 2.5 mM MgCl2

dNTPs: dATP, dCTP, dGTP and dTTP

EvaGreen® dye

ROX reference dye

Bovine serum albumine (BSA) to enhance qPCR reaction

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 an extremely stable dye and has been shown to be non-mutagenic and non-cytotoxic. EvaGreen® is compatible with all common real-time PCR cyclers – simply select the standard settings for SYBR® Green or FAM!

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.

What is the difference between EvaGreen® and SYBR® Green?

Both dyes are DNA intercalating agents that are used to stain dsDNA.

EvaGreen® dye is spectrally similar to FAM or SYBR® Green I, which means that no change in optical settings is required for using an EvaGreen®-based qPCR mix.

EvaGreen® was reported to have several benefits over SYBR® Green I:

  • EvaGreen® dye has less background than SYBR® Green I due to its novel “release-on-demand” DNA-binding mechanism,
  • EvaGreen® is less inhibitory in the PCR reaction than SYBR® Green I, enabling the use of higher dye concentration for maximal signal and better high-resolution DNA melt analysis,
  • EvaGreen® is non-mutagenic, non-cytotoxic and environmentally safe,
  • EvaGreen® is very stable under PCR condition and during routine storage and handling, whereas SYBR® Green I is known to degrade following multiple freeze-thaw cycles and under PCR conditions.
What is the difference between EvaGreen® and SYBR® Green?

Both dyes are DNA intercalating agents that are used to stain dsDNA.

EvaGreen® dye is spectrally similar to FAM or SYBR® Green I, which means that no change in optical settings is required for using an EvaGreen®-based qPCR mix.

EvaGreen® was reported to have several benefits over SYBR® Green I:

  • EvaGreen® dye has less background than SYBR® Green I due to its novel “release-on-demand” DNA-binding mechanism,
  • EvaGreen® is less inhibitory in the PCR reaction than SYBR® Green I, enabling the use of higher dye concentration for maximal signal and better high-resolution DNA melt analysis,
  • EvaGreen® is non-mutagenic, non-cytotoxic and environmentally safe,
  • EvaGreen® is very stable under PCR condition and during routine storage and handling, whereas SYBR® Green I is known to degrade following multiple freeze-thaw cycles and under PCR conditions.
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

Assessment of growth suppression in apple production with replant soils

U. Cavael et al. / Ecol. Indic. / 2020

FoxH1 represses miR-430 during early embryonic development of zebrafish via non-canonical regulation

P. Fischer et al. / BMC Biol. / 2019

Identification of llama KRTAP7-1 and KRTAP8-1 fiber genes and polymorphism screening

M. S. Daverio et al. / Small Rumin. Res. / 2019
read more

A Combination of Real-Time PCR and High-Resolution Melting Analysis to Detect and Identify CpGV Genotypes Involved in Type I Resistance

A. Hinsberger et al. / Viruses / 2019

Informativeness of Single Nucleotide Polymorphisms and Relationships among Onion Populations from Important World Production Regions

M. J. Havey and F. Ghavami / J. Am. Soc. Hortic. Sci. / 2018

L-quebrachitol promotes the proliferation, differentiation, and mineralization of MC3T3-E1 cells: Involvement of the BMP-2/RUnx2/MAPK/Wnt/β-catenin signaling pathway

T. Yodthong et al. / Molecules / 2018

Insights into malaria transmission among Anopheles funestus mosquitoes, Kenya

E. O. Ogola et al. / Parasit. Vectors / 2018

Identification of selected genetic polymorphisms in polycystic ovary syndrome in Sri Lankan women using low cost genotyping techniques

U. Branavan et al. / PLoS One / 2018

Could the interaction between LMX1B and PAX2 influence the severity of renal symptoms?

S. Negrisolo et al. / Eur. J. Hum. Genet. / 2018

Effects of Lactobacillus acidophilus and Bifidobacterium bifidum Probiotics on the Expression of MicroRNAs 135b, 26b, 18a and 155, and Their Involving Genes in Mice Colon Cancer

Z. Heydari et al. / Probiotics and Antimicrobial Proteins / 2018

Study of Ras/MAPK pathway gene variants in Chilean patients with Cryptorchidism

F. Rodríguez et al. / Andrology / 2018

High-resolution melt analysis without DNA extraction affords rapid genotype resolution and species identification

P. F. Rugman-Jones and R. Stouthamer / Mol. Ecol. Resour. / 2017
Storage & Shipping
Storage

Routine storage: -20 oC

Temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of the product

Shipping

At room temperature