DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

Sidore, A. M.; Plesa, C.; Samson, J. A.; Kosuri, S.

doi:10.1101/740977

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bioRxiv · 10.1101/740977

DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

Sidore, A. M.·Plesa, C.·Samson, J. A.·Kosuri, S.

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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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Sidore, A. M., Plesa, C., Samson, J. A., Kosuri, S.. 2019-08-20. DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions. https://doi.org/10.1101/740977

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