Table 2

Summary of different approaches utilized for each genetic situations.

Genetic situation

HRM solutions


No common SNP in the amplicon

Direct HRM analysis


Only one common SNP in the amplicon

Sometimes it is possible to distinguish directly the three groups (Figure 1)

Otherwise, spike-in or unlabelled probe is needed (Figure 2)


Two SNPs in the amplicon, one is common the other is rare

Spike-in when possible (Figure 3); otherwise, use unlabeled probe for the common SNP, and perform mutation scanning for the rare one (Figure 4)


Two or more common SNPs in the amplicon

Use an unlabeled probe for each SNP (Figure 5)

Two SNPs in two different amplicons

When Tm of 2 amplicons is different, it is possible to perform multiplex PCR (Figure 6)


Garritano et al. BMC Genetics 2009 10:5   doi:10.1186/1471-2156-10-5

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