Virtual terminator nucleotides for next-generation DNA sequencing
| Autor: | Philip R. Buzby, Suhaib Siddiqi, Peter McInerney, Kathleen E. Steinmann, Edyta Krzymanska-Olejnik, Jayson Bowers, John F. Thompson, Doron Lipson, Subramanian Marappan, J. William Efcavitch, Judith Mitchell, Marie Causey, Mirna Jarosz, Atanu Roy, Adam R. Platt, Eric Beer, Geoffrey M Lowman, Li Kung |
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| Rok vydání: | 2009 |
| Předmět: |
Chromosomes
Artificial Bacterial Computational biology Biochemistry Sensitivity and Specificity DNA sequencing Article Substrate Specificity 03 medical and health sciences 0302 clinical medicine Dogs Sequencing by hybridization Animals Nucleotide Computer Simulation Molecular Biology Polymerase Chromatography High Pressure Liquid 030304 developmental biology chemistry.chemical_classification 0303 health sciences Bacterial artificial chromosome biology Nucleotides Active site Cell Biology DNA Sequence Analysis DNA Molecular biology Sequencing by ligation Terminator (genetics) chemistry 030220 oncology & carcinogenesis biology.protein Biotechnology |
| Zdroj: | Nature methods |
| ISSN: | 1548-7105 |
| Popis: | Nucleotide analogs modified with a free 3′ hydroxyl, maintaining the interactions at the polymerase active site, and a cleavable linker, attaching a fluorescent dye and an inhibitor, are efficient at reading homopolymer runs in a single-molecule sequencing reaction. We synthesized reversible terminators with tethered inhibitors for next-generation sequencing. These were efficiently incorporated with high fidelity while preventing incorporation of additional nucleotides, and we used them to sequence canine bacterial artificial chromosomes in a single-molecule system that provided even coverage for over 99% of the region sequenced. This single-molecule approach generated high-quality sequence data without the need for target amplification and thus avoided concomitant biases. |
| Databáze: | OpenAIRE |
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