'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
Bang D
中科院分区:
生物学2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

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我们开发了一种高度可扩展的“鸟枪”DNA合成技术,利用微芯片寡核苷酸,鸟枪组装和下一代测序技术。将靶向青霉素生物合成基因簇的微芯片寡核苷酸池组装成许多随机片段,并用20 bp简并条形码引物对标记。通过高通量DNA测序鉴定了一组最佳的无错误片段,使用条形码序列选择性扩增,并成功组装成靶基因簇。
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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