Gene synthesis by circular assembly amplification

Duhee Bang, George M. Church

Research output: Contribution to journalArticlepeer-review

61 Citations (Scopus)

Abstract

Here we report the development of a gene-synthesis technology, circular assembly amplification. In this approach, we first constructed exonuclease-resistant circular DNA via simultaneous ligation of oligonucleotides. Exonuclease- and subsequent mismatch cleaving endonuclease-mediated degradation of the resulting ligation mixture eliminated error-rich products, thereby substantially improving gene-synthesis quality. We used this method to construct genes encoding a small thermostable DNA polymerase, a highly repetitive DNA sequence and large (>4 kb) constructs.

Original languageEnglish
Pages (from-to)37-39
Number of pages3
JournalNature Methods
Volume5
Issue number1
DOIs
Publication statusPublished - 2008 Jan

Bibliographical note

Funding Information:
We thank M. Umbarger, M. Price and J. Aach for critical comments on the manuscript; F. Issacs for comments on experimental design; an anonymous reviewer’s suggestion to test repetitive DNA sequence synthesis. We acknowledge funding from US Department of Energy for GTL Center support. D.B is a Damon Runyon Fellow supported by the Damon Runyon Cancer Research Foundation (DRG#1911-06).

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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