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Vol 51(2017) N 3 p. 474-482; DOI 10.1134/S0026893317030025 Full Text

A.V. Chudinov1*, Y.Y. Kiseleva2, V.E. Kuznetsov1, V.E. Shershov1, M.A. Spitsyn1, T.O. Guseinov1, S.A. Lapa1, E.N. Timofeev1, A.I. Archakov2, A.V. Lisitsa2, S.P. Radko2, A.S. Zasedatelev1

Enzymatic synthesis of high-modified DNA

1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
2Orekhovich Institute of Biomedical Chemistry, Moscow, 119121 Russia

*chud@eimb.ru
Received - 2016-05-10; Accepted - 2016-07-11

Here, we describe the synthesis and purification of six deoxyuridine triphosphate derivatives that contain protein-like functional groups and alkene linkers of various lengths. Using KOD XL and Deep Vent polymerases, these derivatives have been incorporated into single-stranded DNA, achieving a high degree of DNA modification. These polymerases are able to utilize highly modified DNA strands as templates for synthesizing unmodified DNA. The synthesized deoxyuridine triphosphate derivatives are promising as substrates for producing modified aptamers to various target proteins using, e.g., the systematic evolution of ligands by exponential enrichment (SELEX) methodology.

modified nucleotides, deoxyuridine, primer extension, SELEX



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