'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 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.
Department of Chemistry, Yonsei University, Shinchon 134, Seoul 120749, Korea.
Hwangbeom
H
Kim
Hyojun
H
Han
Jinwoo
J
Ahn
Joongoo
J
Lee
Namjin
N
Cho
Hoon
H
Jang
Hyoki
H
Kim
Sunghoon
S
Kwon
Duhee
D
Bang
D004247:DNA / Q000096:biosynthesis*; D059014:High-Throughput Nucleotide Sequencing; D020411:Oligonucleotide Array Sequence Analysis; D010406:Penicillins / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D058615:Synthetic Biology
D010406:Penicillins; D004247:DNA