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https://hdl.handle.net/1822/35361
Título: | Transformation of Escherichia coli JM109 using pUC19 by the Yoshida effect |
Autor(es): | Mendes, Gabriel Vieira, Pedro S. Lanceros-Méndez, S. Kluskens, Leon Mota, M. |
Palavras-chave: | Yoshida effect Transformation Sepiolite E. coli |
Data: | Ago-2015 |
Editora: | Elsevier B.V. |
Revista: | Journal of Microbiological Methods |
Citação: | Mendes, Gabriel Pinto; Vieira, Pedro S.; Lanceros-Mendez, S.; Kluskens, Leon; Mota, M., Transformation of Escherichia coli JM109 using pUC19 by the Yoshida effect. Journal of Microbiological Methods, 115, 1-5, 2015 |
Resumo(s): | Transformation of non-competent Escherichia coli JM109 was accomplished using pUC19 as donor plasmid and sepiolite as the acicular material to promote cell piercing via application of friction with a polystyrene stick or a magnetic bar on the surface of a hydrogel containing agar. An automatic spreading setup was built with a conventional stirring plate and compared to manual spreading. Several parameters were optimized, namely, the agar content of the hydrogel (2%), concentration of cells (OD=1.3 corresponding to 1.4x109 bacterial cells/mL), concentration of sepiolite (0.01%), manual versus mechanical spreading (automatic spreading more consistent) and spreading time (30 sec). Efficiency values up to 4.1 x 104 CFU/µg pUC19 were obtained. The method proved to be suitable for a rapid and low cost transformation of non-competent E. coli JM109, where higher values of efficiency do not need to be attained. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/35361 |
DOI: | 10.1016/j.mimet.2015.05.012 |
ISSN: | 0167-7012 |
e-ISSN: | 0167-7012 |
Versão da editora: | http://www.sciencedirect.com/science/article/pii/S0167701215001517 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: | CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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document_21044_1.pdf | 1,01 MB | Adobe PDF | Ver/Abrir |