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Open Access Publications from the University of California

DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions.

  • Author(s): Sidore, Angus M
  • Plesa, Calin
  • Samson, Joyce A
  • Lubock, Nathan B
  • Kosuri, Sriram
  • et al.

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.

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