A rapid, efficient, and economical inverse polymerase chain reaction-based method for generating a site saturation mutant library.

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Quantitative missense variant effect prediction using large-scale mutagenesis data (2017, neutral): “Variant coverage depended on experimental details such as the protocol used for library generation (e.g., doped oligomer [Matteucci and Heyneker, 1983] versus site saturation mutagenesis [Jain and Varadarajan, 2014]), the number of clones generated, and the sequencing depth.”
Coevolution-based inference of amino acid interactions underlying protein function (2018, methodology): “The region covering the a2 helix is PCR amplified and Truseq barcodes and sequencing adapters (Illumina) are appended in two sequential PCR reactions.”
The inherent mutational tolerance and antigenic evolvability of influenza hemagglutinin (2014, methodology): “However, several PCR-based techniques have recently been developed to introduce random codon mutations into full-length genes (Firnberg and Ostermeier, 2012; Bloom, 2014; Jain and Varadarajan, 2014).”