Economical analysis of saturation mutagenesis experiments

Carlos G. Acevedo-Rocha, Manfred T. Reetz, Yuval Nov

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Article number10654
JournalScientific Reports
StatePublished - 20 Jul 2015

Bibliographical note

Funding Information:
This work was supported by the Max-Planck-Society, the LOEWE Research Cluster SynChemBio from the state of Hessen, Germany; and the Israeli Science Foundation (grant 286/13). C.G.A.R is holder of a SYNMIKRO postdoctoral fellowship awarded by the LOEWE program of the state of Hessen, Germany.

ASJC Scopus subject areas

  • General


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