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...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2025-07-01
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| Series: | BioTechniques |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/07366205.2025.2542012 |
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| author | M. Rey Toleco Mark van der Giezen |
| author_facet | M. Rey Toleco Mark van der Giezen |
| author_sort | M. Rey Toleco |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-5dcc908f0c8d4ff88056f03df54db9bb |
| institution | Kabale University |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-5dcc908f0c8d4ff88056f03df54db9bb2025-08-20T04:02:51ZengTaylor & Francis GroupBioTechniques0736-62051940-98182025-07-011610.1080/07366205.2025.2542012A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kitM. Rey Toleco0Mark van der Giezen1Department of Chemistry, Bioscience, and Environmental Engineering, University of Stavanger, Stavanger, NorwayDepartment of Chemistry, Bioscience, and Environmental Engineering, University of Stavanger, Stavanger, NorwayModular 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.https://www.tandfonline.com/doi/10.1080/07366205.2025.2542012genetic tool developmentmodular cloningNEBuilder® HiFi DNA assemblyBlastocystisNon-model organisms |
| spellingShingle | M. Rey Toleco Mark van der Giezen A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit BioTechniques genetic tool development modular cloning NEBuilder® HiFi DNA assembly Blastocystis Non-model organisms |
| title | A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit |
| title_full | A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit |
| title_fullStr | A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit |
| title_full_unstemmed | A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit |
| title_short | A simple and universal quasi-modular cloning system using NEBuilder® HiFi DNA assembly kit |
| title_sort | simple and universal quasi modular cloning system using nebuilder r hifi dna assembly kit |
| topic | genetic tool development modular cloning NEBuilder® HiFi DNA assembly Blastocystis Non-model organisms |
| url | https://www.tandfonline.com/doi/10.1080/07366205.2025.2542012 |
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