Fig. 4

Single mutation detection in a harsh condition. PNA melting point differences between perfect match target and mutation target containing single SNP, deletion, and insertion were measured by melting point analysis. a Mutation points were distributed in a single amplicon with short gap with other mutation points. b PNA probe fluorescent signals of perfectly matched targets with three PNA probes (left panel) or melting points of PNA probe that was hybridized with single mismatch, deletion, and insertion in a single tube were represented and each signal was listed separately depending on its fluorescence (right panel). c Fluorescence was analyzed after PCR cycles in a closed well reaction. d Sequences of probes and partial targets. Underline, mismatch point; Violet arrow, primer; Red line, Texas-Red; Blue line, FAM; Green line, HEX; PM, perfect match; 1Ins, single insertion; 1Del, single deletion; NTC, non-template control