text: Since the late 1990s product analysis via SYBR Green, other double-strand specific dyes, or probe-based melting curve analysis has become nearly ubiquitous. The probe-based technique is sensitive enough to detect single-nucleotide polymorphisms (SNP) and can distinguish between homozygous wildtype, heterozygous and homozygous mutant alleles by virtue of the dissociation patterns produced. Without probes, amplicon melting (melting and analysis of the entire PCR product) was not generally successful at finding single base variants through melting profiles. With higher resolution instruments and advanced dyes, amplicon melting analysis of one base variants is now possible with several commercially available instruments. For example: Applied Biosystems 7500 Fast System and the 7900HT Fast Real-Time PCR System, Idaho Technologys LightScanner (the first plate-based high resolution melting device), Qiagens Rotor-Gene instruments, and Roche's LightCycler 480 instruments.
  Many research and clinical examples exist in the literature that show the use of melting curve analysis to obviate or complement sequencing efforts, and thus reduce costs.
  While most quantitative PCR machines have the option of melting curve generation and analysis, the level of analysis and software support varies. High Resolution Melt (known as either Hi-Res Melting, or HRM) is the advancement of this general technology and has begun to offer higher sensitivity for SNP detection within an entire dye-stained amplicon. It is less expensive and simpler in design to develop probeless melting curve systems. However, for genotyping applications, where large volumes of samples must be processed, the cost of development may be less important than the total throughput and ease of interpretation, thus favoring probe-based genotyping methods.
  Digital High Resolution Melting (dHRM) is also used in conjunction with digital PCR (dPCR) to improve quantitative power by providing additional information on the melting behavior of the amplified DNA, which can help in distinguishing between different genetic variants and in ensuring the accuracy of the quantification. dHRM is enabled by the use of sensitive DNA-binding dyes and digital PCR instrumentation, which allows for the collection of high-density data points to generate detailed melt profiles. These profiles can be used to identify even subtle differences in nucleic acid sequences, making dHRM a powerful tool for genotyping, mutation scanning, and methylation analysis
  dHRM is an advanced molecular technique used for the analysis of genetic variations, such as single nucleotide polymorphisms (SNPs), mutations, and methylations, by monitoring the melting behavior of double-stranded DNA. It is a post-PCR method that involves the gradual heating of PCR-amplified DNA in the presence of intercalating dyes that fluoresce when bound to double-stranded DNA. As the DNA melts, the fluorescence decreases, and the changes in fluorescence are monitored in real-time with digital PCR system. The resulting melting curves are then analyzed to detect genetic differences based on the melting temperatures of the DNA fragments. 
  The technique has been further advanced by its application on digital microfluidics platforms, which can facilitate the analysis of single-nucleotide polymorphisms (SNPs) with high accuracy and sensitivity. Additionally, massively parallel dHRM has been developed to enable rapid and absolutely quantitative sequence profiling, which can be particularly useful in clinical and industrial settings where accurate quantification of nucleic acids is critical.