Inhibition of non-templated nucleotide addition by DNA polymerases in primer extension using twisted intercalating nucleic acid modified templates
Language English Country England, Great Britain Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
26707394
DOI
10.1016/j.bmcl.2015.12.034
PII: S0960-894X(15)30347-4
Knihovny.cz E-resources
- Keywords
- DNA polymerases, Nucleotide addition, Oligonucleotides, Primer extension, Twisted intercalating nucleic acid,
- MeSH
- Biotin chemistry MeSH
- DNA-Directed DNA Polymerase chemistry genetics MeSH
- DNA chemistry genetics MeSH
- Geobacillus stearothermophilus enzymology MeSH
- Intercalating Agents chemistry MeSH
- Pyrenes chemistry MeSH
- Pyrococcus MeSH
- Thermococcus enzymology MeSH
- Thermus enzymology MeSH
- Triphenylmethyl Compounds chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Biotin MeSH
- DNA-Directed DNA Polymerase MeSH
- DNA MeSH
- Intercalating Agents MeSH
- Pyrenes MeSH
- Triphenylmethyl Compounds MeSH
A simple and elegant method for inhibition of non-templated nucleotide addition by DNA polymerases and for following DNA 3'-heterogeneity in enzymatic DNA synthesis by primer extension (PEX) is described. When template bearing ortho-twisted intercalating nucleic acid (ortho-TINA) at the 5'-end is used, non-templated nucleotide addition is reduced in both the A- and B-family DNA polymerases (KOD XL, KOD (exo-), Bst 2.0, Therminator, Deep Vent (exo-) and Taq). Formation of a single oligonucleotide product was observed with ortho-TINA modified template and KOD XL, KOD (exo-), Bst 2.0, Deep Vent (exo-) and Taq DNA polymerases. This approach can be applied to the synthesis of both unmodified and base-modified oligonucleotides.
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