High Resolution Melting Dyes and probes

The secret of High Resolution Meling (HRM) analysis is to monitor this process happening in real-time. This is achieved by using saturating fluorescent dye. HRM dyes are known as intercalating dyes bind specifically and in high amount (= saturating dye, except SYBR Green) to double-stranded DNA and when they are bound they fluoresce brightly. In the absence of double-stranded DNA they have nothing to bind to and they only fluoresce at a low level (estimations are between 1-4% false positive fluorescence). At the beginning of the HRM analysis, after complete PCR amplification, there is a high level of fluorescence in the sample because of the millions of copies of the amplicon. But as the sample DNA is heated up and the two strands of the DNA denaturate there is no longer any double stranded DNA present and thus fluorescence is reduced =>

Following dyes or probe systems are available:

LCGreen Melting Dyes

LCGreen dyes are specifically designed for high-resolution melting curve analysis to detect DNA sequence variants. The addition of LCGreen dyes increase the melting temperature of DNA by 1-3 °C and may requre adjustment of cycling parameters. The dyes are manufactured exclusively by Idaho Technology and their chemical structures are unique among the scientific and patent literature.
LCGreen dyes are tailored specifically for Hi-Res Melting and have the unique ability to detect heteroduplexes during melting analysis after PCR. Just add the dye to your sample before PCR. LCGreen dyes are extremely stable, do not inhibit PCR, and are "saturation" dyes that can detect multiple PCR products in a mixture during melting analysis.

LCGreen Plus+
RAZOR TrainingLCGreen PLUS is a new member of the dye family tailored for use in melting instruments with 96- or 384-well microtiter plates. It has superb fluorescence intensity and can be used with other fluorescence based PCR detection systems such as the Roche LightCycler®. For optimal performance, the use of a high-resolution melting instrument is required.

LCGreen I
LCGreen I is a dsDNA binding dye used for Hi-Res Melting curve analysis using Idaho Technology’s HR-1™ instrument. This innovative dye is manufactured exclusively by Idaho Technology and is designed specifically for Hi-Res Melting analysis to detect DNA sequence variants (SNPs, Insertions / Deletions).

http://www.idahotech.com/LCGreen/index.html        LC Green Flyer

High-resolution genotyping by amplicon melting analysis using LC Green.
Wittwer CT, Reed GH, Gundry CN, Vandersteen JG, Pryor RJ.
Clin Chem. 2003 49(6 Pt 1): 853-860
BACKGROUND: High-resolution amplicon melting analysis was recently introduced as a closed-tube method for genotyping and mutation scanning (Gundry et al. Clin Chem 2003;49: 396-406). The technique required a fluorescently labeled primer and was limited to the detection of mutations residing in the melting domain of the labeled primer. Our aim was to develop a closed-tube system for genotyping and mutation scanning that did not require labeled oligonucleotides.
METHODS: We studied polymorphisms in the hydroxytryptamine receptor 2A (HTR2A) gene (T102C), beta-globin (hemoglobins S and C) gene, and cystic fibrosis (F508del, F508C, I507del) gene. PCR was performed in the presence of the double-stranded DNA dye
LCGreen, and high-resolution amplicon melting curves were obtained. After fluorescence normalization, temperature adjustment, and/or difference analysis, sequence alterations were distinguished by curve shape and/or position. Heterozygous DNA was identified by the low-temperature melting of heteroduplexes not observed with other dyes commonly used in real-time PCR.
RESULTS: The six common beta-globin genotypes (AA, AS, AC, SS, CC, and SC) were all distinguished in a 110-bp amplicon. The HTR2A single-nucleotide polymorphism was genotyped in a 544-bp fragment that split into two melting domains. Because melting curve
acquisition required only 1-2 min, amplification and analysis were achieved in 10-20 min with rapid cycling conditions. CONCLUSIONS: High-resolution melting analysis of PCR products amplified in the presence of LCGreen can identify both heterozygous and homozygous sequence variants. The technique requires only the usual unlabeled primers and a generic double-stranded DNA dye added before PCR
for amplicon genotyping, and is a promising method for mutation screening.


SYTO 9 green fluorescent nucleic acid stain has been shown to stain live and dead Gram-positive and Gram-negative bacteria, and it is a component of the LIVE/DEAD BacLight Bacterial Viability Kits (L-7007, L-7012, L-13152).

SYTO ® dyes are cell-permeant nucleic acid stains that show a large fluorescence enhancement upon binding nucleic acids. The SYTO dyes can be used to stain RNA and DNA in both live and dead eukaryotic cells, as well as in Gram-positive and Gramneg a tive bacteria. Available as blue-, green-, orange- or redfluorescent dyes, these novel SYTO stains share several important characteristics:

  • Permeability to virtually all cell membranes, including mammalian cells and bacteria
  • High molar absorptivity, with extinction coefficients >50,000 cm-1M-1 at visible absorption maxima
  • Extremely low intrinsic fluorescence, with quantum yields typically <0.01 when not bound to nucleic acids
  • Quantum yields that are typically >0.4 when bound to nucleic acids
SYTO dyes differ from each other in one or more charac ter is tics, including cell permeability, fluorescence enhancement upon binding
nucleic acids, excitation and emission spectra, DNA/RNA selectivity and binding affinity.

Rapid, sensitive, and discriminating identification of Naegleria spp. by real-time PCR and melting-curve analysis.
Robinson BS, Monis PT, Dobson PJ.
Appl Environ Microbiol. 2006 Sep;72(9):5857-63.

Australian Water Quality Centre, Private Mail Bag 3, Salisbury, SA 5108, Australia
The free-living amoeboflagellate genus Naegleria includes one pathogenic and two potentially pathogenic species (Naegleria fowleri, Naegleria italica, and Naegleria australiensis) plus numerous benign organisms. Monitoring of bathing water, water supplies, and cooling systems for these pathogens requires a timely and reliable method for identification, but current DNA sequence-based methods identify only N. fowleri or require full sequencing to identify other species in the genus. A novel closed-tube method for distinguishing thermophilic Naegleria species is presented, using a single primer set and the DNA intercalating dye SYTO9 for real-time PCR and melting-curve analysis of the 5.8S ribosomal DNA gene and flanking noncoding spacers (ITS1, ITS2). Collection of DNA melting data at
close temperature intervals produces highly informative melting curves with one or more recognizable melting peaks, readily distinguished for seven Naegleria species and the related Willaertia magna. Advantages over other methods used to identify these organisms include its comprehensiveness (encompassing all species tested to date), simplicity (no electrophoresis required to verify the product), and sensitivity (unambiguous identification from DNA equivalent to one cell). This approach should be applicable to a wide range of microorganisms of medical importance.

Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and
investigation of the effect of dye concentration on amplification and DNA melting curve analysis.
Monis PT, Giglio S, Saint CP.
Anal Biochem. 2005 340(1): 24-34.
Microbiology Unit, Australian Water Quality Centre, Private Mail Bag 3, Salisbury, SA 5108, Australia.
Following the initial report of the use of SYBR Green I for real-time polymerase chain reaction (PCR) in 1997, little attention has been given to the development of alternative intercalating dyes for this application. This is surprising considering the reported limitations of SYBR Green I, which include limited dye stability, dye-dependent PCR inhibition, and selective detection of amplicons during DNA melting curve analysis of multiplex PCRs. We have tested an alternative to SYBR Green I and report the first detailed evaluation of the
intercalating dye SYTO9. Our findings demonstrate that SYTO9 produces highly reproducible DNA melting curves over a broader range of dye concentrations than does SYBR Green I, is far less inhibitory to PCR than SYBR Green I, and does not appear to selectively detect particular amplicons. The low inhibition and high melting curve reproducibility of SYTO9 means that it can be readily incorporated into a conventional PCR at a broad range of concentrations, allowing closed tube analysis by DNA melting curve analysis. These features simplify the use of intercalating dyes in real-time PCR and the improved reproducibility of DNA melting curve analysis will make SYTO9 useful in a diagnostic context.

The use of new probes and stains for improved assessment of cell viability and
extracellular polymeric substances in Candida albicans biofilms.
Jin Y, Zhang T, Samaranayake YH, Fang HH, Yip HK, Samaranayake LP.
Mycopathologia. 2005 159(3): 353-60.
Division of Oral Biosciences, Faculty Dentistry, The Prince Philip Dental
Hospital, University of Hong Kong, 34 Hospital Road, SAR, China.

Phenotypic and genotypic cell differentiation is considered an important feature that confers enhanced antifungal resistance in candidal biofilms. Particular emphasis has been placed in this context on the viability of biofilm subpopulations, and their heterogeneity with regard to the production of extracellular polymeric substances (EPS). We therefore assessed the utility of two different labeled lectins Erythrina cristagalli (ECA) and Canavalia ensiformis (ConA), for EPS visualization. To evaluate the viability of candidal biofilms, we further studied combination stains, SYTO9 and propidium iodide (PI). The latter combination has been successfully used to assess bacterial, but not fungal, viability although PI alone has been previously used to stain nuclei in fungal cells. Candida albicans biofilms were developed in a rotating disc biofilm reactor and observed in situ using confocal scanning laser microscopy (CSLM). Our data indicate that SYTO9 and PI are reliable vital stains that may be used to investigate C. albicans biofilms. When used together with ConA, the lectin ECA optimized EPS visualization and revealed differential production of this material in mature candidal biofilms. The foregoing probes and stains and the methodology described should help better characterize C. albicans biofilms in terms of cell their viability, and EPS production.


EvaGreen® dye is a green fluorescent nucleic acid dye with features that make the dye useful for several applications including qPCR, high-resolution DNA melt curve analysis (HRM)1, real-time monitoring of thermophilic helicase-dependent amplification (tHDA), routine solution DNA quantification and capillary gel electrophoresis. The DNA-bound dye has excitation and emission spectra very close to those of fluorescein (FAM) or SYBR® dye Green I, making the dye readily compatible with instruments equipped with the 488 nm argon laser or any visible light excitation with wavelength in the region. EvaGreen dye is extremely stable both thermally and hydrolytically, providing convenience during routine handling. The dye is essentially nonfluorescent by itself, but becomes highly fluorescent upon binding to dsDNA. EvaGreen dye is nonmutagenic and noncytotoxic by being completely impermeable to cell membranes, unlike SYBR Green I, which enters cell rapidly and is known to be a powerful mutation-enhancer (Ohta, et el. Mutat. Res. 492, 91(2001).

The unique properties of EvaGreen dye have made it particularly useful in quantitative real-time PCR (qPCR) application. Compared with the widely used SYBR Green I, EvaGreen dye is generally less inhibitory toward PCR and less likely to cause nonspecific amplification. As a result, EvaGreen dye can be used at a much higher dye concentration than SYBR Green I, resulting in more robust PCR signal.

EVAGreen flyer               EVAGreen product information                      
  • Very Little PCR inhibition: Exhibit much less PCR inhibition than SYBR Green I via a smart "release-on-demand" DNA-binding technology.
  • Highly Sensitive:     Low PCR inhibition of the dye permits a higher dye concentration to be used for much greater fluorescent signal and high-resolution melt curve analysis (HRM).
  • Nonmutagenic and noncytotoxic:  Nonmutagenic and noncytotoxic by standard Ames test; completely impermeable to cell membranes (see below).
  • Compatible with Fast PCR protocol:   Minimal interference to PCR makes it possible to significantly shorten the chain extension time.
  • Compatible with multiplex PCR:    No dye migration from amplicon to amplicon when used at the recommended concentration
  • Unsurpassed Thermal Stability, Hydrolytical Stability and Photostability:    No detectable dye decomposition in PCR buffer at 95-100°C for 48 hours; highly stable under either alkaline or acidic condition; withstand repeated freeze-thaw cycles
  • Spectrally similar to SYBR Green I:    Compatible with all major brand qPCR instruments and enzyme systems

Summary of Mutagenic Toxicity Test Results for EvaGreen
Nucleic Acid Detection Technologies
Compiled by Biotium, Inc. from the results of an independent testing service:  Litron Laboratories, Inc., Rochester, NY

DNA quantification using EvaGreen and a real-time PCR instrument.
Weijie Wanga, Kunsong Chena and Changjie XuCorresponding Author Contact Information, a, E-mail The Corresponding Author
aState Agricultural Ministry Laboratory of Horticultural Plant Growth, Development and Biotechnology, Huajiachi Campus, Zhejiang University, Hangzhou 310029, People’s Republic of China
Analytical Biochemistry
Volume 356, Issue 2, 15 September 2006, Pages 303-305
DNA quantification is an important, frequently used technique, and inaccuracies can result in failures with ligation, restriction, polymerase chain reaction (PCR),1 amplified fragment length polymorphism (AFLP), Southern blotting, and other techniques. DNA is most commonly quantified using absorbance at 260 nm, but because of the existence of many impurities, this can be an imprecise measurement and DNA levels can be more than 10 times overestimated in some cases [1]. Quantification by agarose gel electrophoresis with a known amount of standard DNA [1] and [2] can provide more accurate data, but the procedures are complicated, the data often still are not accurate enough, and the technique is impractical for routine or high-throughput DNA quantification [3]. Fluorescence spectroscopy using various DNA intercalating dyes is the most widely accepted technique for accurate DNA quantification [4]. However, if the analysis is to be carried out with a fluorescence spectrophotometer, a relatively large assay volume (e.g., 2 ml) is required [5], and this is impractical for small DNA samples and expensive dyes. Fluorescence can also be measured with a smaller volume of DNA sample using other instruments such as fluorescent microplate readers [6], microplate fluorometers [7] and [8], and transilluminator–microplate–CCD camera systems [9], but the instruments might not be readily available in most molecular biology laboratories.

Characterization of EvaGreen and the implication of its physicochemical properties for qPCR applications.
Mao F, Leung WY, Xin X.
BMC Biotechnol. 2007 7(1): 76
BACKGROUND: EvaGreen (EG) is a newly developed DNA-binding dye that has recently been used in quantitative real-time PCR (qPCR), post-PCR DNA melt curve analysis and several other applications. However, very little is known about the physicochemical properties of the dye and their relevance to the applications, particularly to qPCR and post PCR DNA melt curve analysis. In this paper, we characterized EG along with a widely used qPCR dye, SYBR Green I (SG), for their DNA-binding properties and stability, and compared their performance in qPCR under a variety of conditions. RESULTS: This study systematically compared theDNA binding profiles of the two dyes under different conditions and had these findings: a) EG had a lower binding affinity for both double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) than SG; b) EG showed no apparent preference for either GC- or AT-rich sequence while SG had a slight preference for AT-rich sequence; c) both dyes showed substantially lower affinity toward ssDNA than toward dsDNA and even lower affinity toward shorter ssDNA fragments except that this trend was more pronounced for EG. Our result also demonstrated that EG was stable both under PCR condition and during routine storage and handling. In the comparative qPCR study, both EG and SG exhibited PCR interference when used at high dye concentration, as evident from delayed Ct and/or nonspecific product formation. The problem worsened when the chain extension time was shortened or when the amplicon size was relatively long (>500 bp). However, qPCR using EG tolerated a significantly higher dye concentration, thus permitting a more robust PCR signal as well as a sharper and stronger DNA melt peak. These differences in qPCR performance between the two dyes are believed to be attributable to their differences in DNA binding profiles. CONCLUSION: These findings suggest that an ideal qPCR dye should possess several DNA-binding characteristics, including a "just right" affinity for dsDNA and low or no affinity for ssDNA and short DNA fragments. The favorable DNA-binding profile of EG, coupled with its good stability and instrument-compatibility, should make EG a promising dye for qPCR and related applications.

Capillary electrophoresis of double-stranded DNA fragments using a new fluorescence intercalating dye EvaGreen.
Sang F, Ren J.
J Sep Sci. 2006 29(9): 1275-1280.
College of Chemistry and Chemical Engineering, Shanghai Jiaotong University, Shanghai, P. R. China.

EvaGreen is a new DNA intercalating dye successfully used in quantitative real-time PCR. In the present work, we firstly apply EvaGreen to the analysis of dsDNA by CE with LIF detection. Comparisons of EvaGreen dye with the commonly used dyes SYBR Green I and SYBR Gold were preformed in dsDNA analysis by CE. The linear range of dsDNA using EvaGreen was slightly wider than that using SYBR Gold and SYBR Green I, and the detection limits of dsDNA were not significantly different for the three dyes. Good separations of dsDNA fragments were obtained using the three dyes. Reproducibility of migration time and the peak area of dsDNA fragments with EvaGreen were better than those for SYBR Green I and SYBR Gold. The RSD values were 0.24-0.27% for migration time and 3.45-7.59% for peak area within the same day, 1.35-1.63% for migration time and 6.72-12.05% for peak area for three days. Our data demonstrated that EvaGreen is well suited for the dsDNA analysis by CE with LIF detection.

SYBR Green

HRM with SYBR Green (by Corbett Life Science)

So-called “new generation saturation dyes”, specifically LCGreen® and LCGreen Plus® (Idaho Technology), are promoted by some as essential for successful HRM analysis (Reed, Kent & Wittwer, 2007) although other dyes such as EvaGreen™ (Biotium Inc.) and SYTO®9 (Invitrogen) have been used with similar success (Krypuy et al 2006; Jeffery et al 2007; Wojdacz & Dobrovic 2007).

Moreover, the use of SYBR® Green 1 (SYBR) for HRM is actively discouraged by some authors (Wittwer et al 2003; Reed, Kent & Wittwer, 2007). Liew et al (2004) state that SYBR can only be used for HRM after substantial modification of the protocol (including the use of GC-clamps, triple primers, and allele-specific PCR). By contrast, we and others (Price et al 2007; Pornprasert et al 2008) have found SYBR to be a very successful dye for HRM analysis that does not require any protocol modifications. We unhesitatingly recommend its use with a Rotor-Gene 6000.

Prejudice against SYBR stems from early evidence collected on instrumentation that was not ideal for the task. Original assertions were based on experiments done with a pre-HRM era LightCycler™ capillary-based real-time analyzer (Roche Molecular Systems) using templates that included a low mass DNA size ladder (Wittwer et al 2003; Liew et al 2004). The reason SYBR was less successful than LCGreen was “not entirely clear” at the time, but a “dye redistribution” hypothesis was suggested (Wittwer et al 2003). According to this hypothesis, SYBR dye releases from low-temperature duplexes during melting and dynamically re-intercalates into neighboring duplexes that melt at higher temperature. This mechanism became the basis for the “dye saturation model” (Wittwer et al 2003, Liew et al 2004). According to this model, if sufficiently high concentrations of dye are used such that all binding positions on the DNA are occupied (i.e. saturated), then dye redistribution effects are minimized and greater melting curve resolution can be achieved. So-called “saturating dyes” were defined as those that can be used at concentrations sufficiently high to saturate all DNA binding sites without inhibiting the PCR. This model formed the basis for IP protection and a patent by Witter et al and Idaho Technology.

In spite of the saturation model, mounting evidence clearly shows that it is not valid on the Rotor-Gene 6000 HRM instrument. This is true for a range of dyes, including SYBR (manuscript in preparation). In fact, SYBR used at standard non-saturating concentrations is highly suitable for HRM analysis, as illustrated below for the detection of a Class 4 SNP. We therefore urge users not to dismiss the use of SYBR for HRM.

The reason saturating dye levels are not required for HRM on the Rotor-Gene 6000 (when apparently required on other instrument systems) is not clear. We have limited experience with competing instruments, however it must be noted that the centrifugal rotary format employed by the Rotor-Gene is distinctly different to other HRM instrumentation. Importantly, the Rotor-Gene has 25–50 times the thermal precision of other instruments and the shortest, most sensitive, and most uniform optical path. It also averages multiple readings for each data point reported at each discrete programmed thermal setpoint in a HRM. Surprisingly, Reed et al (2007) claim that the Rotor-Gene can only “approach high-resolution data quality by melting at slower rates”. Ironically, it may be that the slightly slower and more deliberate thermal stepping used by the Rotor-Gene is partly why it achieves superior HRM results without the need for a “saturating dye”.

Genotyping a Class 4 SNP by high resolution melt (HRM) using SYBR Green I
Alister Kwok, Brant Bassam, and Valin Reja,   August 2007

SensiMix HRM     (Quantace)
SensiMix HRM has been designed for High Resolution Melt (HRM) analysis on the Rotor-Gene 6000. To learn more about the applications of HRM and to see some data from our mix, please click here.
  •     HRM optimised mix with a separate vial of EvaGreen dye.
  •     Ultra-high sensitivity: detects class 4 (A/T) SNP mutations.
  •     Comes in 250, 500 and 2000 (25µl) reaction packs.
  •     HRM™ demo kit containing this mix is available to Corbett Life Science Distributors.

Chromofy  for qPCR & HRM

Biocenter, Odinsgatan 28, 41103 Goteborg, Sweden


Chromofy is a monomeric asymmetric cyanine dye, developed by TATAA Biocenter for use in qPCR applications. The dye has absorbance and emission wavelengths that are suitable for the FAM/SYBR channel on most common real-time PCR instruments. When binding to dsDNA Chromofy shows a very strong fluorescence increase.

download flyer

Chromofy™ in High-resolution melting

High-resolution melting is a post-PCR analysis that discriminates different samples on the basis of their melting temperature. Chromofy and HRM can be used for genotyping, either with ro without unlabelled probes and for methylation analysis. Methylation analysis is performed after bisulfite treatment of the DNA samples which converts unmethylated cytosines to uracils while methylated cytosines remain intact. During subsequent PCR the uracils will be substituted for thymines giving rise to a Tm difference between methylated and un-methylated samples, which is seen as varying heights of the plateu in the normalized HRM data. Using Chromofy your assay can be sensitive down to 1 % methylated DNA in un-methylated background.

download Chromofy user manual

BEBO for qPCR  &  HRM

Combining sequence-specific probes and DNA binding dyes in real-time PCR for specific nucleic acid quantification and melting curve analysis
Kristina Lind1,2, Anders Ståhlberg2,3, Neven Zoric2, and Mikael Kubista1,2
1 Chalmers University of Technology, Gothenburg, 2 TATAA Biocenter, Gothenburg, and 3 Lund University, Lund, Sweden
BioTechniques 40:315-319 (March 2006)
Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has the advantage that it only detects the tar-geted product, while the nonspecific dye has the advantage that melting curve analysis can be performed after completed amplification, which reveals what kind of products have been formed. Here we present a new strategy based on combining a sequence-specific probe and a nonspecific dye, BOXTO, in the same reaction, to take the advantage of both chemistries. We show that BOXTO can be used together with both TaqMan® probes and locked nucleic acid (LNA) probes without interfering with the PCR. The probe signal reflect formation of target product, while melting curve analysis of the BOXTO signal reveals primer-dimer formation and the presence of any other anomalous products.

BEBO is an unsymmetric cyanine dye developed by TATAA Biocenter for use in qPCR applications.
The dye has absorbance and emission wavelengths that can be detected on the FAM channel on most common real-time PCR platforms, and shows a strong fluorescence increase when bound to dsDNA. BEBO can be used as an unspecific dye for real-time PCR applications or other applications where staining of dsDNA is wanted.

A new minor groove binding asymmetric cyanine reporter dye for real-time PCR
Martin Bengtsson, H. Jonas Karlsson, Gunnar Westman and Mikael Kubista*
Department of Chemistry and Bioscience, Chalmers University of Technology 41296 Goteborg and
Biocenter, Odinsgatan 28, 41103 Goteborg, Sweden
Nucleic Acids Research, 2003, Vol. 31, No. 8 e45

The minor groove binding asymmetric cyanine dye 4-[(3-methyl-6- (benzothiazol-2-yl)- 2,3-dihydro- (benzo-1,3-thiazole) -2-methylidene)]- 1-methyl-pyridinium iodide (BEBO) is tested as sequence nonspeciÆc label in real-time PCR. The Fluorescence intensity of BEBO increases upon binding to double-stranded DNA allowing emission to be measured at the end of the elongation phase in the PCR cycle. BEBO concentrations between 0.1 and 0.4 mM generated sufÆcient Øuorescence signal
without inhibiting the PCR. A comparison with the commonly used reporter dye SYBR Green I shows that the two dyes behave similarly in all important aspects.

HRM assays with raZor probe
Pre-validated HRM assays    (by Primer Design)     
How does the service work ?
"On demand" HRM assays for any variations in any DNA sequence.  Simply give us the details of your variation of interest and our expert team will professionally design the best possible primers that cater for your individual requirements.  We then scientifically validate the assay on biologically derived gDNA in our own laboratory and package them for you at optimal concentrations for HRM analysis.  The assays arrive with you in less than three weeks and no further optimisation by you is required.

What is an HRM raZor probe?
Custom designed HRM assays that come with a Razor probe are the highest specification HRM assay available.  An especially modified DNA probe targets the site of the SNP or DNA variation of interest.  HRM analysis of the probe/target melting adds huge power the quality of data achieved.  We guarantee the SNP typing power of these assays.