Synthesis and cell-free cloning of DNA libraries using programmable microfluidics
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 pro...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
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Oxford University Press
2016
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| Online Access: | https://eprints.nottingham.ac.uk/34900/ |
| _version_ | 1848794957911949312 |
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| author | Ben Yehezkel, Tuval Rival, Arnaud Raz, Ofir Cohen, Rafael Marx, Zipora Cámara, Miguel Dubern, Jean-Frédéric Koch, Birgit Heeb, Stephan Krasnogor, Natalio Delattre, Cyril Shapiro, Ehud |
| author_facet | Ben Yehezkel, Tuval Rival, Arnaud Raz, Ofir Cohen, Rafael Marx, Zipora Cámara, Miguel Dubern, Jean-Frédéric Koch, Birgit Heeb, Stephan Krasnogor, Natalio Delattre, Cyril Shapiro, Ehud |
| author_sort | Ben Yehezkel, Tuval |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:24:27Z |
| format | Article |
| id | nottingham-34900 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:24:27Z |
| publishDate | 2016 |
| publisher | Oxford University Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-349002020-05-04T17:34:46Z https://eprints.nottingham.ac.uk/34900/ Synthesis and cell-free cloning of DNA libraries using programmable microfluidics Ben Yehezkel, Tuval Rival, Arnaud Raz, Ofir Cohen, Rafael Marx, Zipora Cámara, Miguel Dubern, Jean-Frédéric Koch, Birgit Heeb, Stephan Krasnogor, Natalio Delattre, Cyril Shapiro, Ehud 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. Oxford University Press 2016-02-29 Article PeerReviewed Ben Yehezkel, Tuval, Rival, Arnaud, Raz, Ofir, Cohen, Rafael, Marx, Zipora, Cámara, Miguel, Dubern, Jean-Frédéric, Koch, Birgit, Heeb, Stephan, Krasnogor, Natalio, Delattre, Cyril and Shapiro, Ehud (2016) Synthesis and cell-free cloning of DNA libraries using programmable microfluidics. Nucleic Acids Research, 44 (4). e35/1-e35/12. ISSN 1362-4962 http://nar.oxfordjournals.org/content/44/4/e35 doi:10.1093/nar/gkv1087 doi:10.1093/nar/gkv1087 |
| spellingShingle | Ben Yehezkel, Tuval Rival, Arnaud Raz, Ofir Cohen, Rafael Marx, Zipora Cámara, Miguel Dubern, Jean-Frédéric Koch, Birgit Heeb, Stephan Krasnogor, Natalio Delattre, Cyril Shapiro, Ehud Synthesis and cell-free cloning of DNA libraries using programmable microfluidics |
| title | Synthesis and cell-free cloning of DNA libraries using programmable microfluidics |
| title_full | Synthesis and cell-free cloning of DNA libraries using programmable microfluidics |
| title_fullStr | Synthesis and cell-free cloning of DNA libraries using programmable microfluidics |
| title_full_unstemmed | Synthesis and cell-free cloning of DNA libraries using programmable microfluidics |
| title_short | Synthesis and cell-free cloning of DNA libraries using programmable microfluidics |
| title_sort | synthesis and cell-free cloning of dna libraries using programmable microfluidics |
| url | https://eprints.nottingham.ac.uk/34900/ https://eprints.nottingham.ac.uk/34900/ https://eprints.nottingham.ac.uk/34900/ |