Nature Methods
- 5, 37 - 39 (2008)
Published online: 25 November 2007; | doi:10.1038/nmeth1136
Gene synthesis by circular assembly amplificationDuhee Bang & George M Church
Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA.
Correspondence should be addressed to Duhee Bang dbang@genetics.med.harvard.edu or George M Church http://arep.med.harvard.edu/gmc/email.html. 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.
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