Our laboratory develops simple methods for analyzing DNA. In the early 1990s we introduced rapid-cycle PCR techniques for DNA amplification in 10-15 min. In the mid-1990s, we adapted flow cytometry optics to thermal cycling for real-time monitoring of PCR, resulting in the commercial “LightCycler”. As part of this development, SYBR Green I, hybridization probes, and melting analysis were first applied to real-time PCR. Since the early 2000s, we have focused on melting analysis as a DNA analysis method that does not require any separations, electrophoresis, or covalently-labeled oligonucleotide probes. Only two PCR primers and a dye are needed. DNA melting is a fundamental property of DNA and if followed closely, can provide much more information than previously imagined. We recently developed high-resolution melting methods with saturation dyes to provide rapid, inexpensive methods for mutation scanning (identification of heterozygotes), genotyping and sequence matching. The combination of rapid cycle PCR and high resolution melting analysis enables meaningful results in less than 15 min. We are currently working on web applications that will further simplify the analysis of DNA, such as uMelt (dna.utah.edu) for multiple melting domain predictions of PCR products.
- Real-time PCR
- Polymerase Chain Reaction
- Molecular Diagnostic Techniques
- High resolution melting analysis
- Molecular Genetics
- Flow Cytometry
- DNA Mutational Analysis
- Real-time PCR, high resolution melting, stopped flow fluorescence, time resolved fluorescence. Contact: Carl Wittwer , 581-4737 , 5C124.
- English, fluent.