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Molecules 2015, 20(8), 13591-13602; doi:10.3390/molecules200813591

Synthesis and Enzymatic Incorporation of Modified Deoxyuridine Triphosphates

Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC V6T1Z1, Canada
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Author to whom correspondence should be addressed.
Academic Editor: Ramon Eritja
Received: 2 June 2015 / Revised: 16 July 2015 / Accepted: 20 July 2015 / Published: 24 July 2015
(This article belongs to the Special Issue Frontiers in Nucleic Acid Chemistry)
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Abstract

To expand the chemical functionality of DNAzymes and aptamers, several new modified deoxyuridine triphosphates have been synthesized. An important precursor that enables this aim is 5-aminomethyl dUTP, whereby the pendent amine serves as a handle for further synthetic functionalization. Five functional groups were conjugated to 5-aminomethyl dUTP. Incorporation assays were performed on several templates that demand 2–5 sequential incorporation events using several commercially available DNA polymerases. It was found that Vent (exo-) DNA polymerase efficiently incorporates all five modified dUTPs. In addition, all nucleoside triphosphates were capable of supporting a double-stranded exponential PCR amplification. Modified PCR amplicons were PCR amplified into unmodified DNA and sequenced to verify that genetic information was conserved through incorporation, amplification, and reamplification. Overall these modified dUTPs represent new candidate substrates for use in selections using modified nucleotide libraries. View Full-Text
Keywords: modified nucleoside triphosphates; Vent (exo-) DNA polymerase; incorporation modified nucleoside triphosphates; Vent (exo-) DNA polymerase; incorporation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Liu, E.; Lam, C.H.; Perrin, D.M. Synthesis and Enzymatic Incorporation of Modified Deoxyuridine Triphosphates. Molecules 2015, 20, 13591-13602.

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