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

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

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 f...

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Veröffentlicht in:Nucleic acids research 2012-10, Vol.40 (18), p.e140-e140
Hauptverfasser: Kim, Hwangbeom, Han, Hyojun, Ahn, Jinwoo, Lee, Joongoo, Cho, Namjin, Jang, Hoon, Kim, Hyoki, Kwon, Sunghoon, Bang, Duhee
Format: Artikel
Sprache:eng
Schlagworte:
DNA - biosynthesis
DNA sequencing
Gene clusters
High-Throughput Nucleotide Sequencing
Methods Online
microchips
Oligonucleotide Array Sequence Analysis
Oligonucleotides
Penicillin
Penicillins - biosynthesis
Polymerase Chain Reaction
Primers
Synthetic Biology
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Beschreibung
Zusammenfassung: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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gks546