1. The Synthesis of Designer DNA
- Author
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Colette J. Whitfield, Eimer Tuite, Rachel C. Little, and Andrew R. Pike
- Subjects
biology ,010405 organic chemistry ,Computational biology ,010402 general chemistry ,01 natural sciences ,0104 chemical sciences ,Nucleobase ,chemistry.chemical_compound ,chemistry ,Reannealing ,biology.protein ,DNA ,Polymerase ,Sequence (medicine) - Abstract
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.
- Published
- 2018
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