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

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.

SUBMITTER: Kim H

PROVIDER: S-EPMC3467036 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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

Kim Hwangbeom H Han Hyojun H Ahn Jinwoo J Lee Joongoo J Cho Namjin N Jang Hoon H Kim Hyoki H Kwon Sunghoon S Bang Duhee D

Nucleic acids research 20120616 18

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 ...[more]

PMID: 22705793

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