To provide initial proof of principle for unknown mutation scanning using s-RT-MELT we utilized cell lines and tumor samples containing sequencing-identified mutations at several positions of p53 exon 8. Figure 2A depicts dHPLC chromatograms of the products obtained using a sample containing a p53 exon 8 G4A mutation or a wild-type sample. The standard Surveyor TM -dHPLC approach (28) was first employed to identify the mutation following PCR amplification of exon 8 from genomic DNA. The resulting dHPLC traces contain a single product for the wild-type and two products for the mutation-containing sequences (Figure 2A , curves 1 and 2, respectively). Next, s-RT-MELT was used to screen the same p53 exon 8 sequence. Following PCR amplification Simplex or Multiplex PCR amplification of one or more exons