Enzymatic Assembly of DNA Molecules Up to Several Hundred Kilobases

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2009 Apr 14

PMID 19363495

Citations 4697

Authors

Daniel G Gibson

Lei Young

Ray-Yuan Chuang

J Craig Venter

Clyde A Hutchison 3rd

Hamilton O Smith

Affiliations

Soon will be listed here.

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.

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