We developed a highly scalable 'shotgun' DNA synthesis technology by utilizing microchip oligonucleotides, shotgun assembly and next-generation sequencing technology. A pool of microchip oligonucleotides targeting a penicillin biosynthetic gene cluster were assembled into numerous random fragments, and tagged with 20 bp degenerate barcode primer pairs. An optimal set of error-free fragments were identified by high-throughput DNA sequencing, selectively amplified using the barcode sequences, and successfully assembled into the target gene cluster.
Bibliographical noteFunding Information:
This work was supported by the Intelligent Synthetic Biology Center of the Global Frontier Project funded by the Ministry of Education, Science and Technology, Korea [2011-0031956]. Hwangbeom Kim was supported by a National Junior Research Fellowship, National Research Foundation of Korea [2012-002388]. Funding for open access charge: Ministry of Education, Science and Technology [2011-0031956].
All Science Journal Classification (ASJC) codes