Parallel gene synthesis in a microfluidic device
The ability to synthesize custom de novo DNA constructs rapidly, accurately and inexpensively is highly desired by researchers, as synthetic genes and longer DNA constructs are enabling to numerous powerful applications in both traditional molecular biology and the emerging field of synthetic biolog...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Oxford University Press (OUP)
2012
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| Online Access: | http://hdl.handle.net/1721.1/70996 https://orcid.org/0000-0002-1332-3197 |
| _version_ | 1811086474350690304 |
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| author | Kong, David S. Carr, Peter A., Sr. Chen, Lu Zhang, Shuguang Jacobson, Joseph |
| author2 | Lincoln Laboratory |
| author_facet | Lincoln Laboratory Kong, David S. Carr, Peter A., Sr. Chen, Lu Zhang, Shuguang Jacobson, Joseph |
| author_sort | Kong, David S. |
| collection | MIT |
| description | The ability to synthesize custom de novo DNA constructs rapidly, accurately and inexpensively is highly desired by researchers, as synthetic genes and longer DNA constructs are enabling to numerous powerful applications in both traditional molecular biology and the emerging field of synthetic biology. However, the current cost of de novo synthesis—driven largely by reagent and handling costs—is a significant barrier to the widespread availability of such technology. In this work, we demonstrate, to our knowledge, the first gene synthesis in a microfluidic environment. The use of microfluidic technology greatly reduces reaction volumes and the corresponding reagent and handling costs. Additionally, microfluidic technology enables large numbers of complex reactions to be performed in parallel. Here, we report the fabrication of a multi-chamber microfluidic device and its use in carrying out the syntheses of several DNA constructs. Genes up to 1 kb in length were synthesized in parallel at minute starting oligonucleotide concentrations (10–25 nM) in four 500 nl reactors. Such volumes are one to two orders of magnitude lower than those utilized in conventional gene synthesis. The identity of all target genes was verified by sequencing, and the resultant error rate was determined to be 1 per 560 bases. |
| first_indexed | 2024-09-23T13:26:30Z |
| format | Article |
| id | mit-1721.1/70996 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:26:30Z |
| publishDate | 2012 |
| publisher | Oxford University Press (OUP) |
| record_format | dspace |
| spelling | mit-1721.1/709962022-09-28T14:13:55Z Parallel gene synthesis in a microfluidic device Kong, David S. Carr, Peter A., Sr. Chen, Lu Zhang, Shuguang Jacobson, Joseph Lincoln Laboratory Massachusetts Institute of Technology. Center for Biomedical Engineering Massachusetts Institute of Technology. Center for Bits and Atoms Massachusetts Institute of Technology. Department of Chemical Engineering Massachusetts Institute of Technology. Media Laboratory Jacobson, Joseph Kong, David S. Carr, Peter A., Sr. Chen, Lu Zhang, Shuguang Jacobson, Joseph The ability to synthesize custom de novo DNA constructs rapidly, accurately and inexpensively is highly desired by researchers, as synthetic genes and longer DNA constructs are enabling to numerous powerful applications in both traditional molecular biology and the emerging field of synthetic biology. However, the current cost of de novo synthesis—driven largely by reagent and handling costs—is a significant barrier to the widespread availability of such technology. In this work, we demonstrate, to our knowledge, the first gene synthesis in a microfluidic environment. The use of microfluidic technology greatly reduces reaction volumes and the corresponding reagent and handling costs. Additionally, microfluidic technology enables large numbers of complex reactions to be performed in parallel. Here, we report the fabrication of a multi-chamber microfluidic device and its use in carrying out the syntheses of several DNA constructs. Genes up to 1 kb in length were synthesized in parallel at minute starting oligonucleotide concentrations (10–25 nM) in four 500 nl reactors. Such volumes are one to two orders of magnitude lower than those utilized in conventional gene synthesis. The identity of all target genes was verified by sequencing, and the resultant error rate was determined to be 1 per 560 bases. Massachusetts Institute of Technology. Center for Bits and Atoms National Science Foundation (U.S.) (CBA grant CCR-0122419) 2012-06-01T19:15:09Z 2012-06-01T19:15:09Z 2007-04 2007-01 Article http://purl.org/eprint/type/JournalArticle 0305-1048 1362-4962 http://hdl.handle.net/1721.1/70996 Kong, D. S. et al. “Parallel Gene Synthesis in a Microfluidic Device.” Nucleic Acids Research 35.8 (2007): e61–e61. Web. 1 June 2012. https://orcid.org/0000-0002-1332-3197 en_US http://dx.doi.org/10.1093/nar/gkm121 Nucleic Acids Research Creative Commons Attribution Non-Commercial http://creativecommons.org/licenses/by-nc/2.5 application/pdf Oxford University Press (OUP) Oxford |
| spellingShingle | Kong, David S. Carr, Peter A., Sr. Chen, Lu Zhang, Shuguang Jacobson, Joseph Parallel gene synthesis in a microfluidic device |
| title | Parallel gene synthesis in a microfluidic device |
| title_full | Parallel gene synthesis in a microfluidic device |
| title_fullStr | Parallel gene synthesis in a microfluidic device |
| title_full_unstemmed | Parallel gene synthesis in a microfluidic device |
| title_short | Parallel gene synthesis in a microfluidic device |
| title_sort | parallel gene synthesis in a microfluidic device |
| url | http://hdl.handle.net/1721.1/70996 https://orcid.org/0000-0002-1332-3197 |
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