Self-assembly cloning: a rapid construction method for recombinant molecules from multiple fragments
| Autor: | Akira Matsumoto, Taichi Q. Itoh |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 62, Iss 1, p xxi (2017) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/000114507 |
| Popis: | Protocol Summary Enzyme-free cloning (EFC) can rapidly produce an in-frame fusion gene with multiple fragments. To practically apply EFC, we investigated the extent and sequence of complementary staggered overhangs necessary to direct self-assembly of multiple fragments as well as a size limitation of the constructed DNA molecule. Six-base pair overhangs with 50% GC content were sufficient to direct self-assembly. A functional plasmid that exceeded 10 kb, which includes an in-frame fusion domain, was efficiently constructed from four PCR fragments in one step by our improved method. |
| Databáze: | Directory of Open Access Journals |
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