HRM analysis can also be used to detect methylation differences between genes; this is known as methylation-specific high resolution melting (MS-HRM) analysis.
High Resolution Melting (HRM)
High Resolution Melting (HRM) analysis is a relatively new, post-PCR analysis method used to identify variations in nucleic acid sequences. The method is based on detecting small differences in PCR melting (dissociation) curves. It is enabled by improved dsDNA-binding dyes used in conjunction with real-time PCR instrumentation that has precise temperature ramp control and advanced data capture capabilities. Data are analyzed and manipulated using software designed specifically for HRM analysis.
Advantages of HRM
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Identify More New Variants, Quickly and Accurately
|High Resolution Melting (HRM) analysis is an alternative to dHPLC sequencing screening of new gene variants. The HRM Software is now available on the Applied Biosystems ViiA™ 7, 7900HT Fast, 7500 Fast, StepOnePlus™, and StepOne™ Real-Time PCR Systems. The StepOne™ (48-well) and StepOnePlus™ (96-well) Real-Time PCR Systems are remarkably simple systems designed with a user-friendly, yet powerful, interface for researches of all experience levels. The 7500 Fast Real-Time PCR System delivers precise results with fast thermal cycling in a standard 96-well format. Achieve high-throughput HRM analysis with the 384-well ViiA™ 7 and 7900HT, the gold standard high-throughput systems. The AB HRM application does not require temperature shifting, which results in a greater likelihood of identifying new homozygous mutations than methods that require temperature shifting.||
Figure 1: Difference plot generate on another plate based real-time HRM system.
The ability to easily identify new variants is key for successful HRM applications. An HRM system from Competitor R (Fig 1) was only able to distinguish them in 10% of the population.
By eliminating the temperature shift step, the Applied Biosystems HRM solution (Fig 2) was able to clearly distinguish homozygous variant samples from homozygous wild type samples in 97.5% of the population.
All genotypes were auto-called by the respective software packages and were not altered by the operator. Class 1 SNP (A/G), multiple technical replicates of nine DNA samples representing three genotypes: homozygous wildtype (G/G), homozygous mutant (A/A) and heterozygous (A/G).
- High Resolution Melt (HRM) Software, an Applied Biosystems™ Analysis Module on Thermo Fisher Cloud for most Applied Biosystems™ Real-Time PCR Instruments
- High Resolution Melt (HRM) Software v2.0 (Applied Biosystems™ 7500 Fast System, StepOne™, StepOnePlus™)
- High Resolution Melt (HRM) Software v3.0, 1 license (Applied Biosystems™ 7500 Fast System, StepOne™, StepOnePlus™)
- High Resolution Melt (HRM) Software v3.0, 10 licenses (Applied Biosystems™ 7500 Fast System, StepOne™, StepOnePlus™)
- High Resolution Melt (HRM) Software v3.1, 1 license (Applied Biosystems™ QuantStudio 3, QuantStudio 5, StepOnePlus, 7500 Fast Only)
- High Resolution Melt (HRM) Software v3.1, 10 licenses (Applied Biosystems™ QuantStudio 3, QuantStudio 5, StepOnePlus, 7500 Fast Only)
Download a FREE 30-day trial of High Resolution Melt Software v3.0.1.
MeltDoctor™ High Resolution Melt Reagents
High Resolution Melt Analysis for Mutation Scanning Prior to Sequencing
Methylation Analysis Using Methylation-Sensitive HRM and DNA Sequencing
High Resolution Melting for Genotyping Applications
High-resolution melt for genotyping in Jatropha plants
For Research Use Only. Not for use in diagnostic procedures.