{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["38(8)"],"submitter":["Blake WJ"],"pubmed_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 isolation. Importantly, construction via PSA is sequence-independent, and does not require primary sequence modification (e.g. the addition or removal of restriction sites). The utility of PSA is demonstrated in the construction of a completely synthetic 91-kb chromosome arm from Saccharomyces cerevisiae."],"journal":["Nucleic acids research"],"pagination":["2594-602"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC2860126"],"repository":["biostudies-literature"],"pubmed_title":["Pairwise selection assembly for sequence-independent construction of long-length DNA."],"pmcid":["PMC2860126"],"pubmed_authors":["Chapman BA","Lee ME","Lippow SM","Blake WJ","Baynes BM","Zindal A"],"additional_accession":[]},"is_claimable":false,"name":"Pairwise selection assembly for sequence-independent construction of long-length DNA.","description":"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 isolation. Importantly, construction via PSA is sequence-independent, and does not require primary sequence modification (e.g. the addition or removal of restriction sites). The utility of PSA is demonstrated in the construction of a completely synthetic 91-kb chromosome arm from Saccharomyces cerevisiae.","dates":{"release":"2010-01-01T00:00:00Z","publication":"2010 May","modification":"2025-04-19T06:31:54.964Z","creation":"2019-03-27T00:30:22Z"},"accession":"S-EPMC2860126","cross_references":{"pubmed":["20194119"],"doi":["10.1093/nar/gkq123"]}}