Abstract
Gene synthesis is an invaluable technique in synthetic and molecular biology for synthesis of artificial genes, operons, and even genomes. In many cases the traditional methods for obtaining functional DNA sequences through cloning are not applicable due to the novelty of genetic material. Here, we describe the simple and economical DNA synthesis method based on USERâ„¢ technology. The method consists of (1) synthesis of building blocks up to 500 bp; (2) assembly of genes up to 3 kb; (3) error correction reassembly; and (4) assembly of operons up to 15 kb if needed.
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Acknowledgments
The authors are grateful to Peter Weigele for valuable suggestions. We also thank Laurie Mazzola for DNA sequencing and John Buswell for oligonucleotide synthesis. We are especially thankful for the environment and support provided by Don Comb.
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Vaisvila, R., Bitinaite, J. (2013). Gene Synthesis by Assembly of Deoxyuridine-Containing Oligonucleotides. In: Samuelson, J. (eds) Enzyme Engineering. Methods in Molecular Biology, vol 978. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-293-3_12
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DOI: https://doi.org/10.1007/978-1-62703-293-3_12
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-292-6
Online ISBN: 978-1-62703-293-3
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