Abstract
Massive Mutagenesis® is a proprietary library creation method that enables the fast generation of high-quality genetic libraries. Starting from a single gene on a plasmid and hundreds to thousands of oligonucleotides, a one-step single-strand circular amplification method creates random combinations of several site-directed substitutions, insertions, or deletions. Libraries of up to a billion such variants have been routinely generated. Sequencing those variants demonstrated lower biases than alternative approaches such as error-prone PCR. Screening and selecting them has yielded improved biocatalysts, therapeutic proteins, and antibodies.
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Acknowledgments
Massive Mutagenesis® is a patented technology of Biométhodes, an industrial biotechnology company based near Paris, France. Please contact Biométhodes regarding licensing conditions and partnership opportunities (www.biomethodes.com). The author thanks Ida Swarczewskaja for conceiving and drawing the figure.
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Sylvestre, J. (2010). Massive Mutagenesis®: High-Throughput Combinatorial Site-Directed Mutagenesis. In: Braman, J. (eds) In Vitro Mutagenesis Protocols. Methods in Molecular Biology, vol 634. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-652-8_17
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DOI: https://doi.org/10.1007/978-1-60761-652-8_17
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