The goal of this project is to solve major problems of DNA sequencing chemistry by developing advanced reagents and protocols that are compatible with automated procedures. The project is based upon our remarkable discovery that the majority of sequencing artifacts can be eliminated by incubating our proprietary HyperStable enzyme with template DNA. In Phase I we have demonstrated the feasibility of solving certain deficiencies of cycle sequencing reactions allowing the successful sequencing of unreadable samples. Also, ThermoFidelase I has enable direct automated sequencing of bacterial genomic DNA without the need for cloning or PCR amplification. It works with non-clonable and non-PCRable regions. This breakthrough has major impact on the finishing strategy of large-scale projects and comparative studies of pathogens. During Phase II we will expand our Fidelase collection, develop new topological and enzymatic methods to control DNA structure during thermocycling reaction, optimize protocols and being to develop solutions for the most difficult samples and the direct sequencing of complex eukaryotic genomes. The outcome of this project will have major impact, both on the successful completion of Human Genome Project and advanced strategies in nucleic acid based technologies.
ThermoFidelase will impact sequencing projects on multiple levels. It provides unique options to sequence genomic DNA and the most difficult samples, facilitate the logistics of major genomics projects and streamline problem management. The rapid commercialization of ThermoFidelase is dictated by the explosive growth of DNA sequencing in research and industry, the simplicity of its implementation, and the heightened interest from users and new technology developers.
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| Polushin, N; Malykh, A; Malykh, O et al. (2001) 2'-modified oligonucleotides from methoxyoxalamido and succinimido precursors: synthesis, properties, and applications. Nucleosides Nucleotides Nucleic Acids 20:507-14 |
