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Accurate High-throughput Gene Synthesis Using Programmable DNA Microfluidic Microchips

J. Tian, H. Gong, N. Sheng, X. Zhou, E. Gulari, X. Gao and G.M. Church
Harvard Medical School/Genetics, US

Keywords:
DNA microchip, gene synthesis

Abstract:
Testing the deluge of hypotheses flowing from genomics and systems biology demands accurate and cost-effective gene and genome synthesis. Here we describe a microchip-based technology for multiplex gene synthesis. Pools of thousands of ''construction'' oligonucleotides and tagged complementary ''selection'' oligonucleotides are synthesized on photo-programmable microfluidic chips, released, amplified, and hybridization-selected to reduce synthesis errors. A one-step polymerase assembly multiplexing (PAM) reaction assembles these into multiple genes. This technology enabled the synthesis of all 21 genes that encode the proteins of the E. coli 30S ribosomal subunit and optimization of their translation efficiency in vitro by alteration of codon bias. This is a significant step towards the synthesis of ribosomes in vitro and should have utility for synthetic biology in general.

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