Sequence-independent cloning methods for long DNA fragments applied to synthetic biology
| Autor: | Paulo Henrique Condeixa de França, Regina Maria Miranda Gern, Marcela Savoldi, Thaila Fernanda dos Reis, Nicole Dalonso, Gustavo H. Goldman |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
BIOLOGIA SINTÉTICA Biophysics Gel extraction Computational biology Biochemistry Polymerase Chain Reaction Biotechnological process Ligases 03 medical and health sciences Synthetic biology chemistry.chemical_compound Transformation Genetic Overlap extension polymerase chain reaction Cloning Molecular Molecular Biology Polymerase Simplified methods biology Cell Biology DNA Molecular biology 030104 developmental biology chemistry biology.protein Agarose Synthetic Biology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| ISSN: | 1096-0309 |
| Popis: | Simplified methods to assemble DNA fragments by independent cloning sequence have helped in the progress of synthetic biology, allowing some biotechnological processes to become economically viable by genetic improvement of microorganisms. We compared three methods of assembling six DNA fragments: PCR fusion-based, isothermal NEBuilder and circular polymerase extension cloning (CPEC). Double and triple fusion occurs directly with the PCR products using PCR fusion-based and NEBuilder methods. For multiple fragments the results showed higher efficiency by the CPEC method which allowed assembly of six fragments previously purified by agarose gel extraction, after a sequence of 20 annealing/extension cycles without any primer. |
| Databáze: | OpenAIRE |
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