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DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

View ORCID ProfileAngus M. Sidore, View ORCID ProfileCalin Plesa, Joyce A. Samson, View ORCID ProfileSriram Kosuri
doi: https://doi.org/10.1101/740977
Angus M. Sidore
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, Los Angeles, California, USA
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Calin Plesa
2Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, California, USA
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Joyce A. Samson
2Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, California, USA
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Sriram Kosuri
2Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, California, USA
3UCLA-DOE Institute for Genomics and Proteomics, Molecular Biology Institute, Quantitative and Computational Biology Institute, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, California, USA
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  • For correspondence: sri@ucla.edu
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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 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.

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Posted August 20, 2019.
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DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
Angus M. Sidore, Calin Plesa, Joyce A. Samson, Sriram Kosuri
bioRxiv 740977; doi: https://doi.org/10.1101/740977
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DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
Angus M. Sidore, Calin Plesa, Joyce A. Samson, Sriram Kosuri
bioRxiv 740977; doi: https://doi.org/10.1101/740977

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