A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit

Modular cloning has transformed synthetic biology by enabling rapid assembly of standardized DNA parts, but its effectiveness is often limited to well-characterised model organisms. Here, we expand the utility of the NEBuilder® HiFi DNA Assembly Kit (New England Biolabs) into a quasi-modular system...

Ausführliche Beschreibung

Bibliographische Detailangaben
Veröffentlicht in:BioTechniques. - 1991. - 77(2025), 5-6 vom: 16. Mai, Seite 257-262
1. Verfasser: Toleco, M Rey (VerfasserIn)
Weitere Verfasser: van der Giezen, Mark
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2025
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Blastocystis NEBuilder® HiFi DNA assembly Non-model organisms genetic tool development modular cloning DNA 9007-49-2
Beschreibung
Zusammenfassung:Modular cloning has transformed synthetic biology by enabling rapid assembly of standardized DNA parts, but its effectiveness is often limited to well-characterised model organisms. Here, we expand the utility of the NEBuilder® HiFi DNA Assembly Kit (New England Biolabs) into a quasi-modular system that uses single-stranded oligonucleotide bridges to assemble reusable DNA fragments without the need for restriction enzymes. Using the anaerobic gut eukaryote Blastocystis ST7-B as a test case, we constructed and reassembled vectors with diverse promoter-terminator combinations. These results underscore the versatility of the method and its potential to accelerate genetic tool development in non-model biological systems
Beschreibung:Date Completed 05.09.2025
Date Revised 05.09.2025
published: Print-Electronic
Citation Status MEDLINE
ISSN:1940-9818
DOI:10.1080/07366205.2025.2542012