title: Inhibition of non-templated nucleotide addition by DNA polymerases in primer extension using twisted intercalating nucleic acid modified templates.
abstract: 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.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Science of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, CZ-16610 Prague 6, Czech Republic.
authors.forename: Pedro
authors.initials: P
authors.lastname: Güixens-Gallardo
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Science of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, CZ-16610 Prague 6, Czech Republic; Department of Organic Chemistry, Faculty of Science, Charles University in Prague, Hlavova 8, CZ-12843 Prague 2, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Hocek
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Science of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, CZ-16610 Prague 6, Czech Republic. Electronic address: perlikova@uochb.cas.cz.
authors.forename: Pavla
authors.initials: P
authors.lastname: Perlíková
mesh_terms: D001710:Biotin / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics*; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D007364:Intercalating Agents / Q000737:chemistry*; D011721:Pyrenes / Q000737:chemistry*; D019711:Pyrococcus; D019712:Thermococcus / Q000201:enzymology; D013824:Thermus / Q000201:enzymology; D014320:Triphenylmethyl Compounds / Q000737:chemistry
chemical_list: D007364:Intercalating Agents; D011721:Pyrenes; D014320:Triphenylmethyl Compounds; D001710:Biotin; D004247:DNA; D004259:DNA-Directed DNA Polymerase
keywords: DNA polymerases; Nucleotide addition; Oligonucleotides; Primer extension; Twisted intercalating nucleic acid