'Shotgun DNA synthesis' for the high-throughput construction of large DNA molecules

Hwangbeom Kim; Hyojun Han; Jinwoo Ahn; Joongoo Lee; Namjin Cho; Hoon Jang; Hyoki Kim; Sunghoon Kwon; Duhee Bang

doi:10.1093/nar/gks546

'Shotgun DNA synthesis' for the high-throughput construction of large DNA molecules

Hwangbeom Kim, Hyojun Han, Jinwoo Ahn, Joongoo Lee, Namjin Cho, Hoon Jang, Hyoki Kim, Sunghoon Kwon, Duhee Bang

Department of Chemistry

Research output: Contribution to journal › Article › peer-review

34 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	e140
Journal	Nucleic acids research
Volume	40
Issue number	18
DOIs	https://doi.org/10.1093/nar/gks546
Publication status	Published - 2012 Oct

Bibliographical note

Funding 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

Genetics

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10.1093/nar/gks546

Cite this

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abstract = "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.",

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AU - Cho, Namjin

AU - Jang, Hoon

AU - Kim, Hyoki

AU - Kwon, Sunghoon

AU - Bang, Duhee

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'Shotgun DNA synthesis' for the high-throughput construction of large DNA molecules

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

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