High fidelity TNA synthesis by Therminator polymerase
Open accessAbstract
Therminator DNA polymerase is an efficient DNA-dependent TNA polymerase capable of polymerizing TNA oligomers of at least 80 nt in length. In order for Therminator to be useful for the in vitro selection of functional TNA sequences, its TNA synthesis fidelity must be high enough to preserve successful sequences. We used sequencing to examine the fidelity of Therminator-catalyzed TNA synthesis at different temperatures, incubation times, tNTP ratios and primer/template combinations. TNA synthesis by Therminator exhibits high fidelity under optimal conditions; the observed fidelity is sufficient to allow in vitro selection with TNA libraries of at least 200 nt in length.
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Primer-extension following a mismatch can be much slower than following a Watson-Crick base-pair, and this post-mismatch stalling is significant in non-enzymatic DNA template-copying model systems.
“Primer-extension following a mismatch can be much slower than following a Watson-Crick base-pair. Such post-mismatch stalling was originally noticed in enzymatic reactions [31,32] but is also significant in a non-enzymatic DNA template-copying model system [33].”
From The eightfold path to non-enzymatic RNA replication (2012) · cites this paper as [31]
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- Authors
- Ichida, J. K.
- Affiliations
- Howard Hughes Medical Institute (US)
- Corresponding author
- Ichida, J. K.
- ORCID
- Ichida, J. K.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2005-09-07
- Volume
- 33
- Issue
- 16
- Pages
- 5219-5225
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC1214552
- OpenAlex
- W2160492036
- Cited by
- 104
- References
- 23
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 6 · 2023: 5 · 2022: 5 · 2021: 5 · 2020: 3 · 2019: 1 · 2018: 5 · 2017: 4 · 2016: 8 · 2015: 6 · 2014: 4 · 2013: 7 · 2012: 13
- FWCI
- 1.39
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nucleic Acids Research (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Howard Hughes Medical Institute
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM067986, GM067986, GM053936, R01GM053936)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Physical Sciences › Physics and Astronomy › Astronomy and Astrophysics › Origins and Evolution of Life
- Keywords
- in vitro selection, temperature effects, DNA-dependent RNA polymerase
- MeSH terms
- DNA-Directed DNA Polymerase, Nucleic Acids, Oligonucleotides, Templates, Genetic, Tetroses, Sequence Analysis, DNA
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