The synthesis of designer DNA

Little, RC; Whitfield, CJ; Tuite, EM; Pike, AR

doi:10.1007/978-1-4939-8582-1_2

The synthesis of designer DNA

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

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Abstract

© 2018, Springer Science+Business Media, LLC, part of Springer Nature. The synthesis of designer DNA requires an approach where the user can determine both the sequence and the number of nucleobases. The protocol outlined here describes an enzymatic method for the synthesis of long repeat-sequence DNA. The method utilizes a PCR-based approach; starting with short oligo-seeds, c.a. 20 bp, bearing a minimum of two repeating units of >8 bp sequences. During each heat-cool cycle, the oligo-seeds reanneal imperfectly leaving an overhang, which is then extended by the polymerase. The final length of the DNA is determined by the number of heat-cool cycles performed, reaching up to 20,000 bp after 20 cycles.