title: Deep Mutational Scanning: Library Construction, Functional Selection, and High-Throughput Sequencing.
abstract: Deep mutational scanning is a highly parallel method that uses high-throughput sequencing to track changes in >10(5) protein variants before and after selection to measure the effects of mutations on protein function. Here we outline the stages of a deep mutational scanning experiment, focusing on the construction of libraries of protein sequence variants and the preparation of Illumina sequencing libraries.
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195;
authors.forename: Lea M
authors.initials: LM
authors.lastname: Starita
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington 98195; Department of Medicine, University of Washington, Seattle, Washington 98195; Howard Hughes Medical Institute, Seattle, Washington 98195.
authors.forename: Stanley
authors.initials: S
authors.lastname: Fields
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D005820:Genetic Testing / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics