scholarly journals High-fidelity gene synthesis by retrieval of sequence-verified DNA identified using high-throughput pyrosequencing

2010 ◽  
Vol 28 (12) ◽  
pp. 1291-1294 ◽  
Author(s):  
Mark Matzas ◽  
Peer F Stähler ◽  
Nathalie Kefer ◽  
Nicole Siebelt ◽  
Valesca Boisguérin ◽  
...  
Keyword(s):  
High Throughput ◽  
Gene Synthesis ◽  
High Fidelity

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

2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Subu K Subramanian ◽  
William P Russ ◽  
Rama Ranganathan
Keyword(s):  
Synthetic Biology ◽  
High Throughput ◽  
Dna Sequences ◽  
Gene Synthesis ◽  
Modular Architecture ◽  
Design And Synthesis ◽  
Genetic Components ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Primers ◽  
Microarray Chips

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.

scholarly journals Scalable gene synthesis by selective amplification of DNA pools from high-fidelity microchips

2010 ◽  
Vol 28 (12) ◽  
pp. 1295-1299 ◽  
Author(s):  
Sriram Kosuri ◽  
Nikolai Eroshenko ◽  
Emily M LeProust ◽  
Michael Super ◽  
Jeffrey Way ◽  
...  
Keyword(s):  
Gene Synthesis ◽  
High Fidelity ◽  
Selective Amplification ◽  
Dna Pools

A Novel Platform for High-Throughput Gene Synthesis to Maximize Recombinant Expression in Escherichia coli

2017 ◽  
pp. 113-128 ◽  
Author(s):  
Ana Filipa Sequeira ◽  
Joana L. A. Brás ◽  
Vânia O. Fernandes ◽  
Catarina I. P. D. Guerreiro ◽  
Renaud Vincentelli ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
Recombinant Expression ◽  
Gene Synthesis ◽  
Expression In Escherichia Coli

scholarly journals DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

2020 ◽  
Vol 48 (16) ◽  
pp. e95-e95 ◽  
Author(s):  
Angus M Sidore ◽  
Calin Plesa ◽  
Joyce A Samson ◽  
Nathan B Lubock ◽  
Sriram Kosuri
Keyword(s):  
Error Correction ◽  
Low Cost ◽  
Gene Synthesis ◽  
High Fidelity ◽  
Functional Testing ◽  
Gene Length ◽  
Genetic Elements ◽  
Gene Libraries

Abstract Multiplexed assays allow functional testing of large synthetic libraries of genetic elements, but are limited by the designability, length, fidelity and scale of the input DNA. Here, we improve DropSynth, a low-cost, multiplexed method that builds gene libraries by compartmentalizing and assembling microarray-derived oligonucleotides in vortexed emulsions. By optimizing enzyme choice, adding enzymatic error correction and increasing scale, we show that DropSynth can build thousands of gene-length fragments at >20% fidelity.

Advancing high-throughput gene synthesis technology

2009 ◽  
Vol 5 (7) ◽  
pp. 714 ◽  
Author(s):  
Jingdong Tian ◽  
Kuosheng Ma ◽  
Ishtiaq Saaem
Keyword(s):  
High Throughput ◽  
Gene Synthesis

scholarly journals DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

2019 ◽  
Author(s):  
Angus M. Sidore ◽  
Calin Plesa ◽  
Joyce A. Samson ◽  
Sriram Kosuri
Keyword(s):  
Error Correction ◽  
Low Cost ◽  
Gene Synthesis ◽  
High Fidelity ◽  
Functional Testing ◽  
Gene Length ◽  
Genetic Elements ◽  
Gene Libraries

AbstractMultiplexed assays allow functional testing of large synthetic libraries of genetic elements, but are limited by the designability, length, fidelity and scale of the input DNA. Here we improve DropSynth, a low-cost, multiplexed method which builds gene libraries by compartmentalizing and assembling microarray-derived oligos in vortexed emulsions. By optimizing enzyme choice, adding enzymatic error correction, and increasing scale, we show that DropSynth can build thousands of gene-length fragments at >20% fidelity.

scholarly journals High-throughput, high-fidelity HLA genotyping with deep sequencing

2012 ◽  
Vol 109 (22) ◽  
pp. 8676-8681 ◽  
Author(s):  
C. Wang ◽  
S. Krishnakumar ◽  
J. Wilhelmy ◽  
F. Babrzadeh ◽  
L. Stepanyan ◽  
...  
Keyword(s):  
High Throughput ◽  
Deep Sequencing ◽  
High Fidelity ◽  
Hla Genotyping
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