Augustin, M.A. ; Ankenbauer, W. ; Angerer, B.
Alternative Links zum Volltext:DOIVerlag
| Dokumentenart: | Artikel |
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| Titel eines Journals oder einer Zeitschrift: | Journal of Biotechnology |
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| Verlag: | ELSEVIER SCIENCE BV |
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| Ort der Veröffentlichung: | AMSTERDAM |
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| Band: | 86 |
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| Nummer des Zeitschriftenheftes oder des Kapitels: | 3 |
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| Seitenbereich: | S. 289-301 |
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| Datum: | 2001 |
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| Institutionen: | Biologie und Vorklinische Medizin > Institut für Biophysik und physikalische Biochemie |
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| Identifikationsnummer: | | Wert | Typ |
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| 10.1016/S0168-1656(00)00420-X | DOI |
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| Stichwörter / Keywords: | FLUORESCENT NUCLEOTIDES; CRYSTAL-STRUCTURES; TERNARY COMPLEXES; POLYMERASE-BETA; PROBES; FRAGMENT; COMPILATION; ALIGNMENT; MECHANISM; ANALOGS; single-molecule sequencing; DNA polymerase; fluorescent nucleotides; high-density labeling of DNA |
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| Dewey-Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
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| Status: | Veröffentlicht |
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| Begutachtet: | Ja, diese Version wurde begutachtet |
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| An der Universität Regensburg entstanden: | Ja |
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| Dokumenten-ID: | 73705 |
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Web of Science
Zusammenfassung
The enzymatic incorporation of modified dNTPs into a growing DNA strand has intensively been studied. Modifications were detectable reporter groups such as digoxigenin or biotin, fluorochromes or aliphatic side chains covalently attached to the base. Incorporation efficiencies were determined with several DNA polymerases using linear primer-extension reactions followed by denaturing PAGE as a ...
Zusammenfassung
The enzymatic incorporation of modified dNTPs into a growing DNA strand has intensively been studied. Modifications were detectable reporter groups such as digoxigenin or biotin, fluorochromes or aliphatic side chains covalently attached to the base. Incorporation efficiencies were determined with several DNA polymerases using linear primer-extension reactions followed by denaturing PAGE as a high-resolution detection system. We describe the enzymatic synthesis of DNA consisting of modified nucleotides exclusively. A defined template-primer system allows us to trace incorporation: (1) in up to 18 neighboring positions for several dUTP-derivatives; or (2) in stretches of DNA of up to 40 bases in length with complete substitution of all four natural dNTPs by differently modified counterparts. Synthesized DNA molecules are shown to particularly exhibit dramatically altered physico-chemical properties by contrast with native DNA. These results provide a fundamental data set for probe generation in single-molecule DNA sequencing (SMS). (C) 2001 Elsevier Science B.V. All rights reserved.
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