DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions

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...

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Autores principales: Sidore, Angus M, Plesa, Calin, Samson, Joyce A, Lubock, Nathan B, Kosuri, Sriram
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498354/
https://www.ncbi.nlm.nih.gov/pubmed/32692349
http://dx.doi.org/10.1093/nar/gkaa600
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author Sidore, Angus M
Plesa, Calin
Samson, Joyce A
Lubock, Nathan B
Kosuri, Sriram
author_facet Sidore, Angus M
Plesa, Calin
Samson, Joyce A
Lubock, Nathan B
Kosuri, Sriram
author_sort Sidore, Angus M
collection PubMed
description 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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spelling pubmed-74983542020-09-23 DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions Sidore, Angus M Plesa, Calin Samson, Joyce A Lubock, Nathan B Kosuri, Sriram Nucleic Acids Res Methods Online 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. Oxford University Press 2020-07-21 /pmc/articles/PMC7498354/ /pubmed/32692349 http://dx.doi.org/10.1093/nar/gkaa600 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Sidore, Angus M
Plesa, Calin
Samson, Joyce A
Lubock, Nathan B
Kosuri, Sriram
DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
title DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
title_full DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
title_fullStr DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
title_full_unstemmed DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
title_short DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
title_sort dropsynth 2.0: high-fidelity multiplexed gene synthesis in emulsions
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498354/
https://www.ncbi.nlm.nih.gov/pubmed/32692349
http://dx.doi.org/10.1093/nar/gkaa600
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