'Shotgun DNA synthesis' for the high-throughput construction of large DNA molecules.
'Shotgun DNA synthesis' for the high-throughput construction of large DNA molecules.
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DOI:
10.1093/nar/gks546
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发表时间:
2012-10
影响因子:
14.9
通讯作者:
Bang D
中科院分区:
文献类型:
--
作者:
Kim H;Han H;Ahn J;Lee J;Cho N;Jang H;Kim H;Kwon S;Bang D
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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