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Self-Priming Enzymatic Fabrication of Multiply Modified DNA

Lookup NU author(s): Dr Colette Whitfield, Rachel Little, Professor Bernard Connolly, Dr Eimer Tuite, Dr Andrew Pike

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This is the authors' accepted manuscript of an article that has been published in its final definitive form by Wiley-VCH Verlag, 2018.

For re-use rights please refer to the publisher's terms and conditions.


Abstract

© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim The self-priming synthesis of multiply modified DNA by the extension of repeating unit duplex “oligoseeds” provides a source of versatile DNA. Sterically-demanding nucleotides 5-Br-dUTP, 7-deaza-7-I-dATP, 6-S-dGTP, 5-I-dCTP as well as 5-(octadiynyl)-dCTP were incorporated into two extending oligoseeds; [GATC]5/[GATC]5 and [A4G]4/[CT4]4. The products contained modifications on one or both strands of DNA, demonstrating their recognition by the polymerase as both template (reading) and substrate (writing). Nucleobase modifications that lie in the major groove were reliably read and written by the polymerase during the extension reaction, even when bulky or in contiguous sequences. Repeat sequence DNA over 500 bp long, bearing four different modified units was produced by this method. The number, position and type of modification, as well as the overall length of the DNA can be controlled to yield designer DNA that offers sequence-determined sites for further chemical adaptations, targeted small molecule binding studies, or sensing and sequencing applications.


Publication metadata

Author(s): Whitfield CJ, Little RC, Khan K, Ijiro K, Connolly BA, Tuite EM, Pike AR

Publication type: Article

Publication status: Published

Journal: Chemistry - A European Journal

Year: 2018

Volume: 24

Issue: 57

Pages: 15267-15274

Print publication date: 12/10/2018

Online publication date: 21/06/2018

Acceptance date: 19/06/2018

Date deposited: 01/01/2019

ISSN (print): 0947-6539

ISSN (electronic): 1521-3765

Publisher: Wiley-VCH Verlag

URL: https://doi.org/10.1002/chem.201801976

DOI: 10.1002/chem.201801976

Notes: This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 778001.


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