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Enzymatic Method for the Synthesis of Long DNA Sequences with Multiple Repeat Units

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

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Abstract

A polymerase chain reaction (PCR) derived method for preparing long DNA, consisting of multiple repeat units of one to ten base pairs, is described. Two seeding oligodeoxynucleotides, so-called oligoseeds, which encode the repeat unit and produce a duplex with 5-overhangs, are extended using a thermostable archaeal DNA polymerase. Multiple rounds of heat-cool extension cycles, akin to PCR, rapidly elongate the oligoseed. Twenty cycles produced long DNA with uniformly repeating sequences to over 20kilobases (kb) in length. The polynucleotides prepared include [A](n)/[T](n), [AG](n)/[TC](n), [A(2)G](n)/[T2C](n), [A(3)G](n)/[T3C](n), [A(4)G](n)/[T4C](n), [A(9)G](n)/[T9C](n), [GATC](n)/[CTAG](n), and [ACTGATCAGC](n)/[TGACTAGTCG](n), indicating that the method is extremely flexible with regard to the repeat length and base sequence of the initial oligoseeds. DNA of this length (20kb approximate to 7m) with strictly defined base reiterations should find use in nanomaterial applications.


Publication metadata

Author(s): Whitfield CJ, Turley AT, Tuite EM, Connolly BA, Pike AR

Publication type: Article

Publication status: Published

Journal: Angewandte Chemie International Edition

Year: 2015

Volume: 54

Issue: 31

Pages: 8971-8974

Print publication date: 27/07/2015

Online publication date: 10/06/2015

ISSN (print): 1433-7851

ISSN (electronic): 1521-3773

Publisher: Wiley - VCH Verlag GmbH & Co. KGaA

URL: http://dx.doi.org/10.1002/anie.201502971

DOI: 10.1002/anie.201502971


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