A set of experimentally validated, mutually orthogonal primers for combinatorially specifying genetic components

Subu K. Subramanian, William P. Russ, Rama Ranganathan

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

The design and synthesis of novel genes and deoxyribonucleic acid (DNA) sequences is a central technique in synthetic biology. Current methods of high throughput gene synthesis use pooled oligonucleotides obtained from custom-designed DNA microarray chips, and rely on orthogonal (non-interacting) polymerase chain reaction primers to specifically de-multiplex, by amplification, the precise subset of oligonucleotides necessary to assemble a full length gene. The availability of a large validated set of mutually orthogonal primers is therefore a crucial reagent for high-throughput gene synthesis. Here, we present a set of 166 20-nucleotide primers that are experimentally verified to be non-interacting, capable of specifying 13 695 unique genes. These primers represent a valuable resource to the synthetic biology community for specifying genetic components that can be assembled through a scalable and modular architecture.

Original languageEnglish (US)
Article numberysx008
JournalSynthetic Biology
Volume3
Issue number1
DOIs
StatePublished - Jun 6 2018

Keywords

  • DNA assembly
  • genetic circuit
  • modular genetic engineering
  • orthogonal primers

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomaterials
  • Biomedical Engineering
  • Agricultural and Biological Sciences (miscellaneous)

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