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

ABSTRACT: 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.

SUBMITTER: Kim H 

PROVIDER: S-EPMC3467036 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature