Precise, highly reproducible and fast genotyping of sequence variants through high resolution melting.
Class 4 SNPs are the most challenging to differentiate; having Tm changes typically less than 0.2oC. The three different genotypes of an A>T SNP were analysed on the Rotor-Gene 6000, using SensiFAST HRM Kit. A difference plot of this data illustrates the effect of subtracting an average heterozygous curve from all curves and plotting the results.
SensiFAST HRM Kit has been developed for detailed characterization of samples according to their base composition, length and GC content by high resolution melting. The latest advances in buffer chemistry and enhancers, together with an antibody-mediated hot-start DNA polymerase system, ensure the SensiFAST HRM Kit delivers reproducible, accurate HRM analysis.
SensiFAST HRM Kit contains EvaGreen®, a third generation, saturating fluorescent dye which selectively binds to double-stranded DNA. In contrast to dyes such as SYBR® Green I, EvaGreen can be used at higher concentrations without inhibiting PCR and shows equal binding affinity for GC-rich and AT-rich regions. The combination of our SensiFAST DNA Polymerase, unique buffer system and EvaGreen dye enables amplification and discrimination of even the most challenging sequence differences, such as class IV SNPs without sequence preference. Since it does not require expensive labelled oligonucleotide probes, SensiFAST HRM Kit is also a cost effective alternative to traditional probe based genotyping methods.
| Concentration | 2x |
| Presentation | BIO-32005: 500 x 20 µL Reactions: 5 x 1 mL BIO-32020: 2000 x 20 µL Reactions: 4 x 5 mL |
| Appearance | Clear, colorless solution |
| Hot Start | Antibody mediated |
| Application | Probe-based, qPCR, two-step RT-qPCR |
| Sample type | cDNA, DNA |
| Presentation | 5 vials / 4 vials |
| Storage | -20 °C, avoid exposure to light |
| Mix stability | See outer label |
| Consistency | Genotypes called with ≥90% confidence |
| DNA Contamination | None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. |
| DNase Contamination | No detectable degradation |
Cat. No. Size
BIO-32005 500 Reactions
BIO-32020 2000 Reactions
Dr Simon Baker, Senior Director of R&D, Meridian, London
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