Inhibition of non-templated nucleotide addition by DNA polymerases in primer extension using twisted intercalating nucleic acid modified templates

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• 作者：Pedro Gü ; ixens-Gallardo^a ; Michal Hocek^a ; ^b ; Pavla Perlí ; ková ; ^a ; ^{perlikova@uochb.cas.cz" class="auth_mail" title="E-mail the corresponding author}
• 关键词：DNA polymerases ; Nucleotide addition ; Primer extension ; Oligonucleotides ; Twisted intercalating nucleic acid
• 刊名：Bioorganic & Medicinal Chemistry Letters
• 出版年：2016
• 出版时间：15 January 2016
• 年：2016
• 卷：26
• 期：2
• 页码：288-291
• 全文大小：666 K

文摘

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.