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Nature Research, Scientific Reports, 1(5), 2015

DOI: 10.1038/srep10654

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Economical analysis of saturation mutagenesis experiments

Journal article published in 2015 by Carlos G. Acevedo Rocha ORCID, Manfred T. Reetz, Yuval Nov
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

AbstractSaturation 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.