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Published online 15 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 7 e59
© 2004 Oxford University Press

Two-step total gene synthesis method

Lei Young* and Qihan Dong

Department of Medicine, University of Sydney, Australia

*To whom correspondence should be addressed. Tel: +61 411688392; Fax: +61 295161273; Email: lyoung{at}student.usyd.edu.au

Received January 2, 2004; Revised and Accepted March 19, 2004

In the post-genomic era, the ability to synthesize any arbitrary DNA sequence is increasingly in demand. A bottleneck in current gene synthesis technologies is the associated cost, due primarily to the high cost of oligonucleotides synthesis and post-synthesis sequencing. In the present paper, an improved method for low-cost gene synthesis that combines dual asymmetrical PCR and overlap extension PCR is presented, which enables any DNA sequence to be synthesized error free. Additionally, the method is easily amenable to automation.


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