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Updated: Aug 4, 2026

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
Published on: July 3, 2016
A method for introducing site-specific mutations using oligonucleotide primers and its application to site-saturation
1Laboratoire de Médiateurs Protéiques, INRA Centre de Jouy-en-Josas, France.
Abstract:
Oligonucleotide primer-directed mutagenesis is a useful molecular biological tool, which is invaluable for the study of the structure/function relationships in proteins and for the creation of mutant proteins possessing modified or novel biological activities. Mutagenesis studies in which a site-saturation approach is employed require a high-efficiency mutagenesis procedure, which will generate a population of mutated molecules containing an even distribution of all possible amino acid changes, or a subset thereof. This article describes such a mutagenesis technique and discusses the adaptations that are necessary to perform site-saturation mutagenesis.
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