Abstract
Gene synthesis by chemical methods provides a powerful tool for modifying genes and exploring their structure, expression, and function in the post-genomic era. However, a bottleneck in recent gene synthesis technologies is the high cost of oligonucleotide synthesis and post-synthesis sequencing. Here, we describe a simple, rapid, and low-cost gene synthesis method based on overlap extension PCR (OE-PCR) and the DNAWorks program. This method enables DNA sequences with sizes ranging from 200 bp to 3 kb to be synthesized with few errors, and these errors can be easily corrected by site-directed mutagenesis. Thus, it is amenable to automation for the multiplexed synthesis of different genes and has a potential for high-throughput gene synthesis.
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Acknowledgements
This work was supported by the grant of the National Natural Science Foundation of China (No. 31170117, 31270156), the National Marine Research Special Funds for Public Welfare Projects of China (201205020), the Major Science and Technology Projects of Guangdong Province, China (2011A080403006), the Fundamental Research Fund for the Central Universities of Sun Yat-sen University (No. 11lgpy23), the key grant of Science and Technology Department of Henan Province (112102210385), and Science Foundation for the high-level talent of Nanyang Normal University (zx20110007).
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Li, G., Dong, BX., Liu, YH., Li, CJ., Zhang, LP. (2013). Gene Synthesis Method Based on Overlap Extension PCR and DNAWorks Program. In: Polizzi, K., Kontoravdi, C. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1073. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-625-2_2
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DOI: https://doi.org/10.1007/978-1-62703-625-2_2
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-624-5
Online ISBN: 978-1-62703-625-2
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