Fluorescent detection and isolation of DNA variants using stabilized RecA-coated oligonucleotides

Extract: The information generated by the genome sequencing initiatives is being gathered, cataloged and evaluated. These databases will provide fertile ground for the development of new technologies in the areas of functional genomics and molecular diagnostics. Concurrent use of these data sets, however, presents an ethical dilemma. Genomic sequence data reveals a substantial amount of sequence diversity within a population; therefore, confidentiality must be maintained so that an individual is not exposed to the risk of "genetic" discrimination. In addition, as nucleic acid tests enter this diagnostic arena, the obtaining of a high degree of accurate and reliable data from the assays will become essential for decision making in healthcare. Genotyping methodologies, which aim to identify genetic variants in large populations often rely heavily on the use of PCR to amplify the designated template DNA. By its very nature, PCR requires a denaturation step that produces the substrates for primer annealing and template amplification. In some cases, nucleic acid denaturation and the number of processing steps in the PCR reaction can introduce errors into the template, mistakes that are then amplified logarithmically creating a sequence bias. In addition, specimen handling of PCR samples and/or products provides the basis for a certain degree of user contamination. Considering the importance of producing exact genotypic IDs for individuals and its real life implications, new techniques that reduce the potential for mistakes at any stage of the processes resulting in data for analysis should be pursued vigorously.