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In the 2000s, we developed 2 instruments (the HR-1 and the LightScanner) that set the standard for quality melting analysis, but they are no longer commercially available. The HR-1 used LightCycler capillaries, requiring only 1-2 minutes for melting. The LightScanner used 96- or 384-well plates with a 5 min turnaround, but did not temperature cycle for PCR. These early instruments showed that melting can be performed rapidly – indeed, with microfluidics, SNP typing by melting can occur in seconds, specifically 1-4 seconds across a 30°C temperature range (Pryor et al. 2017 , Myrick et al. 2017). With the right instruments to precisely control and measure fluorescence and temperature, high resolution melting can be performed very quickly.

Convenience is often easier to sell than speed, so commercial instruments today melt at a much slower rate, often requiring 10-90 minutes to achieve the same resolution. Unfortunately, speed helps to discriminate multiple products, including the different genotypes of SNPs. Faster cooling and heating rates support detection of the kinetically-favored heteroduplexes over the equilibrium-favored homoduplexes (Gundry et al. 2003).



Although genotyping can be performed without this speed advantage for heteroduplex detection, genotyping accuracy and sensitivity can improve at faster speeds. There is no strict definition of high resolution melting (often referred to as HRM, HRMA, or Hi-Res melting) and available instruments differ widely in their ability to resolve different genotypes (Herrmann et al. 2006, Herrmann et al. 2007, Herrmann et al. 2007).