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.

• 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