A set of experimentally validated, mutually orthogonal primers for combinatorially specifying genetic components. Issue 1 (18th January 2018)

• Record Type: Journal Article
• Authors: Subramanian, Subu K; Russ, William P; Ranganathan, Rama
• Is Part Of: Synthetic biology. Volume 3:Issue 1(2018)
• Publication Date: 2018-01-18
• Subjects: orthogonal primers -- DNA assembly -- modular genetic engineering -- genetic circuit; Synthetic biology -- Periodicals; 660.6
• DOI: 10.1093/synbio/ysx008
• Languages: English
• ISSNs: 2397-7000

Abstract

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.