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

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.

SUBMITTER: Subramanian SK

PROVIDER: S-EPMC7445780 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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

Subramanian Subu K SK Russ William P WP Ranganathan Rama R

Synthetic biology (Oxford, England) 20180118 1

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 s ...[more]

PMID: 32995509

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