Abstract
We experienced significant problems while developing a PyrosequencingTM (Biotage, Uppsala, Sweden) assay to characterize the rifampin resistance-determining region of Mycobacteriumtuberculosis, a target with high guanosine-cytosine content. This paper describes the successful use of a modified pyrosequencing protocol through partial substitution of deoxyguanosine triphosphate with deoxyinosine triphosphate in the polymerase chain reaction.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacterial Proteins / genetics
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Base Composition
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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DNA, Bacterial / metabolism*
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Deoxyguanosine / metabolism*
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Humans
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Inosine / analogs & derivatives*
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Inosine / metabolism
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Mycobacterium tuberculosis / genetics
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Pathology, Molecular / methods*
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Sequence Analysis, DNA / methods*
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Specimen Handling / methods*
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Sweden
Substances
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ARR-2 protein, Bacteria
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Bacterial Proteins
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DNA, Bacterial
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Inosine
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Deoxyguanosine
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deoxyinosine