Improvement of Low-Density Microelectronic Array Technology to Characterize 14 Mutations/Single-Nucleotide Polymorphisms from Several Human Genes on a Large Scale

peaks, which indicate the presence or absenceof a PCR product, to be strategically placed within thetypical melting curve range of 45–70 °C for hybridizationprobes. Over this melting curve range we feel it should bepossible to clearly distinguish three peaks in a singlechannel. This potentially makes it possible to multiplexsix mutations in two capillaries when both fluorescentchannels are used. This enhances the capabilities of theLightCycler for mutation detection; similar strategiescould also be used for other real-time systems.The results presented here demonstrate the ability tocarry out multiplex mutation detection by use of a com-bination of ARMS PCR and real-time detection. A labora-tory currently using ARMS PCR in a diagnostic settingcan quite easily convert the standard ARMS PCR to areal-time ARMS PCR. This could have a major advantagein time savings and reduce the handling of potentialcarcinogenic ethidium bromide.