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Synthesis and cell-free cloning of DNA libraries using programmable microfluidics

Ben Yehezkel, Tuval; Rival, Arnaud; Raz, Ofir; Cohen, Rafael; Marx, Zipora; C�mara, Miguel; Dubern, Jean-Fr�d�ric; Koch, Birgit; Heeb, Stephan; Krasnogor, Natalio; Delattre, Cyril; Shapiro, Ehud

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Authors

Tuval Ben Yehezkel

Arnaud Rival

Ofir Raz

Rafael Cohen

Zipora Marx

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MIGUEL CAMARA MIGUEL.CAMARA@NOTTINGHAM.AC.UK
Professor of Molecular Microbiology

Jean-Fr�d�ric Dubern

Birgit Koch

Natalio Krasnogor

Cyril Delattre

Ehud Shapiro



Abstract

Microfluidics may revolutionize our ability to write synthetic DNA by addressing several fundamental limitations associated with generating novel genetic constructs. Here we report the first de novo synthesis and cell-free cloning of custom DNA libraries in sub-microliter reaction droplets using programmable digital microfluidics. Specifically, we developed Programmable Order Polymerization (POP), Microfluidic Combinatorial Assembly of DNA (M-CAD) and Microfluidic In-vitro Cloning (MIC) and applied them to de novo synthesis, combinatorial assembly and cellfree cloning of genes, respectively. Proof-of-concept for these methods was demonstrated by programming an autonomous microfluidic system to construct and clone libraries of yeast ribosome binding sites and bacterial Azurine, which were then retrieved in individual droplets and validated. The ability to rapidly and robustly generate designer DNA molecules in an autonomous manner should have wide application in biological research and development.

Citation

Ben Yehezkel, T., Rival, A., Raz, O., Cohen, R., Marx, Z., Cámara, M., …Shapiro, E. (2016). Synthesis and cell-free cloning of DNA libraries using programmable microfluidics. Nucleic Acids Research, 44(4), e35. https://doi.org/10.1093/nar/gkv1087

Journal Article Type Article
Acceptance Date Oct 3, 2015
Online Publication Date Oct 19, 2015
Publication Date Feb 29, 2016
Deposit Date Jul 13, 2016
Publicly Available Date Jul 13, 2016
Journal Nucleic Acids Research
Print ISSN 0305-1048
Electronic ISSN 1362-4962
Publisher Oxford University Press
Peer Reviewed Peer Reviewed
Volume 44
Issue 4
Pages e35
DOI https://doi.org/10.1093/nar/gkv1087
Public URL https://nottingham-repository.worktribe.com/output/774668
Publisher URL http://nar.oxfordjournals.org/content/44/4/e35
Contract Date Jul 13, 2016

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