Figure 3. Multiplex Real time ARMS / High resolution melt PCR.. A: 2.5% ethidium bromide stained agarose gel showing the results of multiplex PCR amplification of (1) unmutated DNA and (2–4) subject DNA with mutations in 3460A, 1177A, and 14484C. The control 221 bp Ins PCR product is present in all lanes with the allele-specific products appearing only in the 3460A (120 bp), 1177A (143 bp), and 14484C (161 bp) subject samples. B: Multiplex real-time melt curves show clear differentiation of the melt peaks for the control (90. 68 °C), 3460A (82.32 °C), 11,778 (80.12 °C), and 14484C (76.32 °C). All samples produce the control products while the presence of a mutation is detected by the additional allele-specific amplification of the mutation-specific products with a melt temperature at the appropriate temperature.