High Resolution Melting (HRM) analysis is a post-PCR technique used to identify genetic variation in nucleic acid sequences. HRM is often used to screen large numbers of potential variant sequences quickly and inexpensively as a precursor to other more laborious techniques such as sequencing. The resulting melt curve profiles can provide valuable information for genotyping studies in addition to many other applications, including mutation screening and methylation analysis.
Applied Biosystems MeltDoctor HRM reagents are specifically formulated to deliver sensitive and consistent results with HRM applications such as SNP analysis, methylation analysis, and mutation scanning.
HRM analysis differs from standard melt curve analysis (with SYBR Green dye, for example) in two ways:
While TaqMan SNP Genotyping Assays are currently the gold standard for accurate and reproducible PCR-based genotyping, HRM is well suited for specific use cases. For example, when inherent sequence limitations prevent you from designing TaqMan probes, or when you are genotyping highly mutable samples where probe specificity may miss some species, HRM may be more successful. Similarly, consider HRM when you are analyzing many SNPs with low sample numbers, or analyzing nucleic acid variation in samples containing possibly unknown SNPs.
Other genotyping studies that can benefit from HRM include:
MeltDoctor HRM Master Mix contains all components needed for HRM-PCR (excluding template and primers). It is formulated for superior HRM performance across a wide range of genomic targets. Unlike some mixes on the market, MeltDoctor HRM Master Mix does not require additional mixing prior to use and was developed and optimized solely for HRM applications. Components of the master mix, provided in a convenient 2X mix, include:
A highly purified DNA polymerase that provides hot-start performance, minimizing nonspecific product formation and enabling reactions to be set up at room temperature
Stabilized form of the fluorescent Invitrogen SYTO 9 double-stranded nucleic acid stain with optical and chemical properties important for high-performance HRM
Minimizes carryover contamination by allowing amplicon degradation by uracil-DNA glycosylase (UDG) in subsequent PCR reactions
Magnesium salts and other buffer components are precisely formulated to obtain optimal HRM results
MeltDoctor HRM Master Mix greatly simplifies your experimental setup as it contains everything you need. Just add your sample and PCR primer pair and you're ready to run your reaction.
The kit contains PCR components and MeltDoctor HRM dye required for high resolution melting analysis. Reagents are supplied unmixed in separate containers so that you can customize your experiments.
This positive control kit contains components to demonstrate and troubleshoot HRM analysis, including forward and reverse PCR primers and three DNA templates representing alleles AA, AB and BB.
Ready-to-use MeltDoctor HRM Calibration Plates contain all the components required for pure dye and HRM calibration, including MeltDoctor HRM Calibration Standard and MeltDoctor HRM Master Mix, reducing the complexity of getting started with HRM experimentation. They provide thermal and optical calibration required for accurate and reproducible discrimination between melt curves.
The MeltDoctor HRM Calibration Standard contains the DNA template and primers for researchers who prefer to prepare their own calibration plate for thermal and optical HRM calibration.
For Research Use Only. Not for use in diagnostic procedures.