Inhibition of non-templated nucleotide addition by DNA polymerases in primer extension using twisted intercalating nucleic acid modified templates
Jazyk angličtina Země Anglie, Velká Británie Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
26707394
DOI
10.1016/j.bmcl.2015.12.034
PII: S0960-894X(15)30347-4
Knihovny.cz E-zdroje
- Klíčová slova
- DNA polymerases, Nucleotide addition, Oligonucleotides, Primer extension, Twisted intercalating nucleic acid,
- MeSH
- biotin chemie MeSH
- DNA-dependentní DNA-polymerasy chemie genetika MeSH
- DNA chemie genetika MeSH
- Geobacillus stearothermophilus enzymologie MeSH
- interkalátory chemie MeSH
- pyreny chemie MeSH
- Pyrococcus MeSH
- Thermococcus enzymologie MeSH
- Thermus enzymologie MeSH
- tritylové sloučeniny chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biotin MeSH
- DNA-dependentní DNA-polymerasy MeSH
- DNA MeSH
- interkalátory MeSH
- pyreny MeSH
- tritylové sloučeniny 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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