Precise sequential DNA ligation on a solid substrate: solid-based rapid sequential ligation of multiple DNA molecules.

ABSTRACT: Ligation, the joining of DNA fragments, is a fundamental procedure in molecular cloning and is indispensable to the production of genetically modified organisms that can be used for basic research, the applied biosciences, or both. Given that many genes cooperate in various pathways, incorporating multiple gene cassettes in tandem in a transgenic DNA construct for the purpose of genetic modification is often necessary when generating organisms that produce multiple foreign gene products. Here, we describe a novel method, designated PRESSO (precise sequential DNA ligation on a solid substrate), for the tandem ligation of multiple DNA fragments. We amplified donor DNA fragments with non-palindromic ends, and ligated the fragment to acceptor DNA fragments on solid beads. After the final donor

SUBMITTER: Takita E 

PROVIDER: S-EPMC3859325 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature