High Resolution Melting (HRM)

High Resolution Melting (HRM) analysis is a relatively new, post-PCR analysis method used to identify variation 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

Low reagent consumption, with little waste: HRM requires only PCR reaction volume (20 µL) for analysis of each sample, eliminating the need for HPLC solvents or DGGE gels
Simple, fast workflow: no additional instrumentation is required after PCR amplification. A high-resolution melt curve can be added to the end of the amplification run and analyzed immediately after completion
Fast optimization: unlike DHPLC, thermal optimization is not required
Low sample consumption: following HRM analysis, the PCR amplicon can be used directly in a Sanger sequencing reaction

We offer a complete guide to performing and optimizing your HRM experiments.
Register and View the HRM Guide.

	Methylation HRM analysis can also be used to detect methylation differences between genes; this is known as methylation-specific high resolution melting (MS-HRM) analysis.		Mutation Scanning High Resolution Melt analysis can be used to scan for mutations in target genes for the identification of variant samples prior to sequencing analysis.
	No Temperature Shift Required Minimal temperature shift of DNA melting due to dye binding.		HRM Complete Product Offering For information on Applied Biosystems solutions for HRM analysis – including instrumentation, software, and reagents – access the “Product Literature” tab at right.