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Literature DB >> 19363495

Enzymatic assembly of DNA molecules up to several hundred kilobases.

Daniel G Gibson¹, Lei Young, Ray-Yuan Chuang, J Craig Venter, Clyde A Hutchison, Hamilton O Smith.

Abstract

We describe an isothermal, single-reaction method for assembling multiple overlapping DNA molecules by the concerted action of a 5' exonuclease, a DNA polymerase and a DNA ligase. First we recessed DNA fragments, yielding single-stranded DNA overhangs that specifically annealed, and then covalently joined them. This assembly method can be used to seamlessly construct synthetic and natural genes, genetic pathways and entire genomes, and could be a useful molecular engineering tool.

Mesh：

Substances：

Year: 2009 PMID： 19363495 DOI： 10.1038/nmeth.1318

Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547

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