The most commonly observed form of human sequence variation is single nucleotide polymorphisms (SNPs), which can affect protein function, proper processing of genes or affect the normal level of gene expression. We propose the development of a novel approach to fluorescence detection with application for high-throughput identification of informative SNPs, which could lead to more accurate diagnosis of inherited disease, better prognosis of risk susceptibilities, or identification of sporadic mutations. The proposed technology is called Pulse-Multiline Excitation or PME. The PME technology has two potential advantages, which could significantly increase fluorescence sensitivity: (1) optimal excitation of all fluorophores in the genomic assay and (2) """"""""color-blind"""""""" detection, which collects considerably more light. This technology differs significantly from the current state-of-the-art DNA sequencing instrumentation, which features single source excitation and color dispersion for DNA sequence identification. To test the feasibility of PME for multi-color fluorescence detection, we propose the construction of a single capillary breadboard, detailed limit of detection experiments to assess sensitivity, and reconstruction experiments for DNA sequencing of SNPs. Successful implementation of the PME technology will have broad application for routine usage in clinical diagnostics, forensics, and general sequencing methodologies and will have the capability, flexibility, and portability of targeted sequence variation assays for a large majority of the population.
| Metzker, Michael L (2010) Sequencing technologies - the next generation. Nat Rev Genet 11:31-46 |
| Metzker, Michael L (2005) Emerging technologies in DNA sequencing. Genome Res 15:1767-76 |
| Lewis, Ernest K; Haaland, Wade C; Nguyen, Freddy et al. (2005) Color-blind fluorescence detection for four-color DNA sequencing. Proc Natl Acad Sci U S A 102:5346-51 |
