Nano Science and Technology Institute
Nanotech 2005 Vol. 1
Nanotech 2005 Vol. 1
Technical Proceedings of the 2005 NSTI Nanotechnology Conference and Trade Show, Volume 1
Chapter 10: Micro and Nano Fluidics Design and Phenomena

Accurate High-throughput Gene Synthesis Using Programmable DNA Microfluidic Microchips

Authors:J. Tian, H. Gong, N. Sheng, X. Zhou, E. Gulari, X. Gao and G.M. Church
Affilation:Harvard Medical School/Genetics, US
Pages:628 - 631
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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