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

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.

SUBMITTER: Sidore AM

PROVIDER: S-EPMC7498354 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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

Sidore Angus M AM Plesa Calin C Samson Joyce A JA Lubock Nathan B NB Kosuri Sriram S

Nucleic acids research 20200901 16

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-lengt ...[more]

PMID: 32692349

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