Pairwise selection assembly for sequence-independent construction of long-length DNA.

ABSTRACT: The engineering of biological components has been facilitated by de novo synthesis of gene-length DNA. Biological engineering at the level of pathways and genomes, however, requires a scalable and cost-effective assembly of DNA molecules that are longer than approximately 10 kb, and this remains a challenge. Here we present the development of pairwise selection assembly (PSA), a process that involves hierarchical construction of long-length DNA through the use of a standard set of components and operations. In PSA, activation tags at the termini of assembly sub-fragments are reused throughout the assembly process to activate vector-encoded selectable markers. Marker activation enables stringent selection for a correctly assembled product in vivo, often obviating the need for clonal isolati

SUBMITTER: Blake WJ 

PROVIDER: S-EPMC2860126 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature