Augustin, M. A. and Ankenbauer, W. and Angerer, B. (2001) Progress towards single-molecule sequencing: enzymatic synthesis of nucleotide-specifically labeled DNA. JOURNAL OF BIOTECHNOLOGY, 86 (3). pp. 289-301. ISSN 0168-1656
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled 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 |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 02 Feb 2022 12:30 |
| Last Modified: | 02 Feb 2022 12:30 |
| URI: | https://pred.uni-regensburg.de/id/eprint/41502 |
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