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Economical analysis of saturation mutagenesis experiments

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Acevedo-Rocha,  Carlos G.
Research Department Reetz, Max-Planck-Institut für Kohlenforschung, Max Planck Society;
Philipps-Universität Marburg, Fachbereich Chemie;
Max Planck Inst Terr Mikrobiol, Prokaryot Small RNA Biol Grp, D-35043 Marburg, Germany;
Univ Marburg, Landes Offens Entwicklung Wissensch Okon Exzellen, D-35032 Marburg, Germany;

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Reetz,  Manfred T.
Research Department Reetz, Max-Planck-Institut für Kohlenforschung, Max Planck Society;
Philipps-Universität Marburg, Fachbereich Chemie;

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Zitation

Acevedo-Rocha, C. G., Reetz, M. T., & Nov, Y. (2015). Economical analysis of saturation mutagenesis experiments. Scientific Reports, 5, 10654. doi:10.1038/srep10654.


Zitierlink: https://hdl.handle.net/11858/00-001M-0000-0029-2288-0
Zusammenfassung
Saturation mutagenesis is a powerful technique for engineering proteins, metabolic pathways and genomes. In spite of its numerous applications, creating high-quality saturation mutagenesis libraries remains a challenge, as various experimental parameters influence in a complex manner the resulting diversity. We explore from the economical perspective various aspects of saturation mutagenesis library preparation: We introduce a cheaper and faster control for assessing library quality based on liquid media; analyze the role of primer purity and supplier in libraries with and without redundancy; compare library quality, yield, randomization efficiency, and annealing bias using traditional and emergent randomization schemes based on mixtures of mutagenic primers; and establish a methodology for choosing the most cost-effective randomization scheme given the screening costs and other experimental parameters. We show that by carefully considering these parameters, laboratory expenses can be significantly reduced.