DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

Angus M Sidore; Calin Plesa; Joyce A Samson; Nathan B Lubock; Sriram Kosuri

doi:10.1093/nar/gkaa600

DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

Nucleic Acids Res. 2020 Sep 18;48(16):e95. doi: 10.1093/nar/gkaa600.

Authors

Angus M Sidore¹, Calin Plesa², Joyce A Samson², Nathan B Lubock², Sriram Kosuri^{2

3}

Affiliations

¹ Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ UCLA-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, CA 90095, USA.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Emulsions / chemistry
Escherichia coli / genetics
Gene Library*
Genes, Synthetic*
Nucleic Acid Amplification Techniques / methods*
Oligonucleotides / genetics*

Substances

Emulsions
Oligonucleotides

Grants and funding

DP2 GM114829/GM/NIGMS NIH HHS/United States