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Cold Spring Harb Protoc. 2015 Aug 3;2015(8):777-80. doi: 10.1101/pdb.prot085225.

Deep Mutational Scanning: Library Construction, Functional Selection, and High-Throughput Sequencing.

Author information

1
Department of Genome Sciences, University of Washington, Seattle, Washington 98195;
2
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.

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.

PMID:
26240405
DOI:
10.1101/pdb.prot085225
[Indexed for MEDLINE]

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