The Sequence Saturation Mutagenesis (SeSaM) method

Abstract

Sequence Saturation Mutagenesis (SeSaM) is a random mutagenesis method developed to overcome the limitations of existing error-prone PCR (epPCR) protocols. SeSaM is advantageous with respect to (1) elimination of mutagenic "hot spots", (2) increase in frequency of subsequent nucleotide substitutions, (3) control over the mutational bias through the utilization of universal base analogs, and, consequently, (4) the prospect of generating transversion-enriched mutant libraries. These advanced features lead to chemically diverse mutant libraries on the protein level, essentially making SeSaM a complementary technology to transition biased epPCR mutagenesis methods.