‘Shotgun DNA synthesis’ for the high-throughput construction of large DNA molecules | Zendy

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

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

Author(s) -

Hwangbeom Kim,

Hyojun Han,

Jinwoo Ahn,

Joongoo Lee,

Namjin Cho,

Hoon Jang,

Hyoki Kim,

Sunghoon Kwon,

Duhee Bang

Publication year - 2012

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gks546

Subject(s) - biology , shotgun sequencing , oligonucleotide , shotgun , dna , dna sequencing , barcode , computational biology , primer (cosmetics) , dna nanoball sequencing , genomic library , genetics , gene , microbiology and biotechnology , base sequence , computer science , chemistry , organic chemistry , operating system

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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