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Updated: Jun 24, 2026

Self-Assembly of Gamma-Modified Peptide Nucleic Acids into Complex Nanostructures in Organic Solvent Mixtures
Published on: June 26, 2020
Enzymatic assembly of DNA molecules up to several hundred kilobases
Daniel G Gibson1, Lei Young, Ray-Yuan Chuang
1The J. Craig Venter Institute, Synthetic Biology Group, Rockville, Maryland, USA. dgibson@jcvi.org
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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