No training required: experimental tests support homology-based DNA assembly as a best practice in synthetic biology

Journal of Biological Engineering

Mads Kærn¹  Shihab Sarwar⁴  Alexandra C Tzahristos⁴  Martin Hanzel²  Nada Elnour³  Vaibhav Gupta³  Alex Power³  Daniel Jedrysiak³  Ian J Roney³  Lioudmila Tepliakova³  Hilary Phenix³  Wilson Lam⁴  Afnan Azizi³ 

[1] Department of Physics, University of Ottawa, MacDonald Hall, Ottawa, K1N 6N5, Ontario, Canada;Biomedical Sciences Program, University of Ottawa, Marion Hall, Ottawa, K1N 6N5, Ontario, Canada;Department of Cellular and Molecular Medicine, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5, Ontario, Canada;Biochemistry Program, University of Ottawa, Gendron Hall, Ontario, K1N 6N5, Canada

关键词: PCR;    iGEM;    Seamless;    Gibson;    Cloning;    DNA assembly;    Synthetic biology;   

Others  :  1213953
DOI  :  10.1186/s13036-015-0006-z

 received in 2015-03-26, accepted in 2015-05-12,  发布年份 2015

【 摘 要 】

The Registry of Standard Biological Parts imposes sequence constraints to enable DNA assembly using restriction enzymes. Alnahhas et al. (Journal of Biological Engineering 2014, 8:28) recently argued that these constraints should be revised because they impose an unnecessary burden on contributors that use homology-based assembly. To add to this debate, we tested four different homology-based methods, and found that students using these methods on their first attempt have a high probability of success. Because of their ease of use and high success rates, we believe that homology-based assembly is a best practice of Synthetic Biology, and recommend that the Registry implement the changes proposed by Alnahhas et al. to better support their use.

【 授权许可】

   
2015 Azizi et al.; licensee BioMed Central.

【 图 表 】

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