Improvement of low-density microelectronic array technology to characterize 14 mutations/single-nucleotide polymorphisms from several human genes on a large scale.

a peak within 1–2 °C of the predicted T m. This allows T m peaks, which indicate the presence or absence of a PCR product, to be strategically placed within the typical melting curve range of 45–70 °C for hybridization probes. Over this melting curve range we feel it should be possible to clearly distinguish three peaks in a single channel. This potentially makes it possible to multiplex six mutations in two capillaries when both fluorescent channels are used. This enhances the capabilities of the LightCycler for mutation detection; similar strategies could also be used for other real-time systems. The results presented here demonstrate the ability to carry out multiplex mutation detection by use of a combination of ARMS PCR and real-time detection. A laboratory currently using ARMS PCR in a diagnostic setting can quite easily convert the standard ARMS PCR to a real-time ARMS PCR. This could have a major advantage in time savings and reduce the handling of potential carcinogenic ethidium bromide. Sensitivity of single-stranded conforma-tion polymorphism and heteroduplex method for mutation detection in the cystic fibrosis gene. Limitations of melting curve analysis using Sybr Green 1—fragment differentiation and mutation detection in the CFTR-gene. Rapid screening for five major cystic fibrosis mutations by melting peak analysis using fluorogenic hybridisation probes. In: Dietmaier W, Wittwer C, Sivasubramanian N, eds. Rapid cycle real-time PCR methods and applications (genetics and oncology). Berlin: Springer, 2002:85–94. 8. Glaab W, Skopek TR. A novel assay for allelic discrimination that combines the fluorogenic 5Ј nuclease polymerase chain reaction (TaqMan ®) and mismatch amplification mutation assay. Spreadsheet software for thermodynamic melting point prediction of oligonucleotide hybridization with and without mis-matches. Application of a thermodynamic nearest-neighbor model to estimate nucleic acid stability and optimize probe design: prediction of melting points of multiple mutations of apolipoprotein B-3500 and factor V with a hybridization probe genotyping assay on the LightCycler. Large-scale human genetic studies require new technologies to genotype several samples with relative ease, high accuracy, and reasonable costs. Among the available approaches, a microelectronic array technology has been developed for DNA hybridization analysis of mutations/ single-nucleotide polymorphisms (SNPs) (1– 4). The mi-croelectronic array system (NanoChip ® Molecular Biology Workstation; Nanogen) produces a defined electric field that allows charged molecules, such as nucleic acids, to be transported to any test site, or pad, on the electronic chip (NanoChip cartridge). Electronic-based molecule addressing can rapidly achieve a high concentration of …