Synthesis of libraries and multi-site mutagenesis using a PCR-derived, dU-containing template.

ABSTRACT: Directed DNA libraries are useful because they focus genetic diversity in the most important regions within a sequence. Ideally, all sequences in such libraries should appear with the same frequency and there should be no significant background from the starting sequence. These properties maximize the number of different sequences that can be screened. Described herein is a method termed SLUPT (Synthesis of Libraries via a dU-containing PCR-derived Template) for generating highly targeted DNA libraries and/or multi-site mutations wherein the altered bases may be widely distributed within a target sequence. This method is highly efficient and modular. Moreover, multiple distinct sites, each with one or more base changes, can be altered in a single reaction. There is very low background from

SUBMITTER: Meinke G 

PROVIDER: S-EPMC8260824 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature