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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| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2025-07-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/07366205.2025.2542012 |
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| Summary: | 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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| ISSN: | 0736-6205 1940-9818 |