Virtual terminator nucleotides for next-generation DNA sequencing

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 sy...

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Veröffentlicht in:	Nature methods 2009-08, Vol.6 (8), p.593-595
Hauptverfasser:	Thompson, John F, Bowers, Jayson, Mitchell, Judith, Beer, Eric, Buzby, Philip R, Causey, Marie, Efcavitch, J William, Jarosz, Mirna, Krzymanska-Olejnik, Edyta, Kung, Li, Lipson, Doron, Lowman, Geoffrey M, Marappan, Subramanian, McInerney, Peter, Platt, Adam, Roy, Atanu, Siddiqi, Suhaib M, Steinmann, Kathleen
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Bioinformatics Biological Microscopy Biological Techniques Biomedical and Life Sciences Biomedical Engineering/Biotechnology brief-communication Chromatography, High Pressure Liquid Chromosomes Chromosomes, Artificial, Bacterial - chemistry Chromosomes, Artificial, Bacterial - genetics Computer Simulation DNA - chemistry DNA polymerase DNA sequencing Dogs Genomics Life Sciences Ligases Methods Molecular biology Nucleotide sequencing Nucleotides Nucleotides - chemistry Nucleotides - genetics Proteomics Scientific method Sensitivity and Specificity Sequence Analysis, DNA - methods Substrate Specificity
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Beschreibung
Zusammenfassung:	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.
ISSN:	1548-7091 1548-7105
DOI:	10.1038/nmeth.1354