2nd July 2024

AccuMelt HRM SuperMix

Accurately characterize genetic variations from novel mutations to rare polymorphisms and more.

AccuMelt HRM SuperMix maximises differences in melt temperature and curve shape, to allow discrimination of DNA sequence differences amongst different samples.  AccuMelt is suitable for a broad range of applications including SNP analysis, mutation scanning, transgene analysis, species identification and DNA methylation analysis.

  • See subtle differences in sequence clearly – robust amplification ensures sufficient yield of products to generate discrete melt curves
  • Accurate genotype calling – comparable or better performance than TaqMan® Genotyping
  • Work with rare or precious samples – large range of template inputs possible
  • Specificity – works with lower Mg2+ concentration than other systems thus enhancing assay accuracy.
AccuMelt HRM SuperMix is a 2x concentrated, hot-start, ready to use reaction cocktail for detection of genetic variations using high resolution melting (HRM) analysis. It includes all required components except for primers and DNA template. HRM is a closed tube, rapid and cost effective procedure for characterization of sequences differences immediately following PCR amplification. It is based on the melting (dissociation) behavior of a PCR product as it transitions from double stranded to single stranded DNA in the presence of SYTO 9 green fluorescent dsDNA binding dye.
AccuStart Taq DNA Polymerase allows for room temperature reaction assembly and storage at +4°C for 6 months.
fig1

Superior Resolution of Genotypes

SNP Genotyping is a useful application for HRM and illustrates the capabilities of AccuMelt HRM SuperMix. Genotypes are readily identified based on unique melting profiles depending on a sample’s sequence. Furthermore, AccuMelt HRM SuperMix gives superior resolution of difficult genotypes when compared to the leading competitor’s mix based on greater Tm differences observed for A -> T transversions.

superior resolution

Comparison to TaqMan genotyping

TaqMan Genotyping has been used successfully in SNP analysis and other allelic discrimination applications. This widely adopted standard in genotyping was used as a benchmark to assess the utility of HRM with our SuperMix. AccuMelt HRM was determined to be just as effective as TaqMan Genotyping in SNP analysis and was even able to call the genotype for a difficult sample which the TaqMan assay could not resolve.

taqman comparison

Robust Amplification

Consistent robust amplification is critical to accuracy in HRM analysis. AccuMelt HRM SuperMix will drive all PCR amplifications to plateau regardless of the quantity of template input. This ensures accurate results regardless of the quantity of DNA available.

robust amplification

Contents

  • 2 x reaction buffer containing optimized concentrations of MgCl2
  • dNTPs (including dUTP)
  • AccuStart Taq DNA Polymerase
  • SYTO® 9 green-fluorescent dye
  • stabilizers
  • Free Mg2+= 0.8mM at 1x final concentration.

Storage conditions

AccuMelt HRM SuperMix is stable for up to 1 year when stored in a constant temperature freezer at 20°C, protected from light. For convenience, it may be stored unfrozen at +2 to +8°C for up to 6 months. After thawing, mix thoroughly by gently vortexing tube contents. Centrifuge briefly to collect contents before using. While repeated freezing and thawing of the product is not recommended, AccuMelt HRM SuperMix demonstrated no loss of performance after 20 cycles of freezing and thawing.

Resources:

Ordering

Cat# 95103-250 250 x 20µl rxns (2 x 1,25 ml)
Cat# 95103-012 1250 x 20µl rxns (10 x 1,25 ml)
Cat# 95103-05K 5000 x 20µl rxns (2 x 25 ml)