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Nucleic Acids Res. 2012 Oct;40(18):e140. doi: 10.1093/nar/gks546. Epub 2012 Jun 16.

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

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1
Department of Chemistry, Yonsei University, Shinchon 134, Seoul 120749, Korea.

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.

PMID:
22705793
PMCID:
PMC3467036
DOI:
10.1093/nar/gks546
[Indexed for MEDLINE]
Free PMC Article
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