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...
| Veröffentlicht in: | BioTechniques. - 1991. - 77(2025), 5-6 vom: 16. Mai, Seite 257-262 |
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| Format: | Online-Aufsatz |
| Sprache: | English |
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2025
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| 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 |
| 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 |
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| 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 |