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Isotachophoresis for rapid transformation of Escherichia coli

Citation

Alves, MN and Nai, YN and Powell, SM and Macka, M and Breadmore, MC, Isotachophoresis for rapid transformation of Escherichia coli, Electrophoresis, 43, (4) pp. 543-547. ISSN 1522-2683 (2022) [Refereed Article]


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DOI: doi:10.1002/elps.202100095

Abstract

A frequent limitation of electroporation (EP) and chemical transformation (CT) are the need of tedious and time-consuming procedures for inducing transformation competence, the substantial number of cells required, and the low transformation yields typically achieved. Here, we show a new and rapid electrokinetic method for transformation of small number of noncompetent Escherichia coli TOP10 cells (2-3 105 ) at room temperature. Escherichia coli TOP10 cells and plasmid DNA are sequentially injected into a 50 μm ID capillary and focused into 11.5 nL by isotachophoresis (ITP) induced by application of high DC voltage (-16 kV). Through ITP, a large excess of plasmid DNA is brought in contact with the cell surface, with the contact time adjusted by application of a counter-pressure (1.3 psi) opposing the ITP movement. The transformation rate was more than 1000-fold higher compared to EP and CT at survival rates greater than 60%.

Item Details

Item Type:Refereed Article
Keywords:molecular biology, microbiology
Research Division:Chemical Sciences
Research Group:Analytical chemistry
Research Field:Separation science
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the biological sciences
UTAS Author:Alves, MN (Ms Monica Alves)
UTAS Author:Powell, SM (Dr Shane Powell)
UTAS Author:Macka, M (Professor Mirek Macka)
UTAS Author:Breadmore, MC (Professor Michael Breadmore)
ID Code:154717
Year Published:2022
Deposited By:Agriculture and Food Systems
Deposited On:2022-12-23
Last Modified:2023-01-10
Downloads:0

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